The RAG2 C terminus suppresses genomic instability
LETTER
doi:10.1038/nature09755
The RAG2C terminus suppresses genomic instability and lymphomagenesis
Ludovic Deriano 1,Julie Chaumeil 1,Marc Coussens 1,Asha Multani 2,YiFan Chou 1,Alexander V.Alekseyenko 3,Sandy Chang 4,Jane A.Skok 1,5&David B.Roth 1
Misrepair of DNA double-strand breaks produced by the V(D)J recombinase (the RAG1/RAG2proteins)at immunoglobulin (Ig)and T cell receptor (Tcr)loci has been implicated in pathogenesis of lymphoid malignancies in humans 1and in mice 2–7.Defects in DNA damage response factors such as ataxia telangiectasia mutated (ATM)protein and combined deficiencies in classical non-homologous end joining and p53predispose to RAG-initiated genomic rearrange-ments and lymphomagenesis 2–11.Although we showed previously that RAG1/RAG2shepherd the broken DNA ends to classical non-homologous end joining for proper repair 12,13,roles for the RAG proteins in preserving genomic stability remain poorly defined.Here we show that the RAG2carboxy (C)terminus,although dis-pensable for recombination 14,15,is critical for maintaining genomic stability.Thymocytes from ‘core ’Rag2homozygotes (Rag2c /c mice)show dramatic disruption of Tcr a /d locus integrity.Furthermore,all Rag2c /c p532/2mice,unlike Rag1c /c p532/2and p532/2animals,rapidly develop thymic lymphomas bearing complex chromosomal translocations,amplifications and deletions involving the Tcr a /d and Igh loci.We also find these features in lymphomas from Atm 2/2mice.We show that,like ATM-deficiency 3,core RAG2severely desta-bilizes the RAG post-cleavage complex.These results reveal a novel genome guardian role for RAG2and suggest that similar ‘end release/end persistence’mechanisms underlie genomic instability and lymphomagenesis in Rag2c /c p532/2and Atm 2/2mice.
RAG mutations can cause specific defects in the joining stage of V(D)J recombination 12,13,16.The ‘dispensable’RAG2C terminus (murine amino acids 1–383)is of particular interest:loss of the RAG2C terminus impairs joining of substrates 17,increases levels of double-strand breaks 17that persist through the cell cycle 18,and increases accessibility of the broken DNA ends to alternative non-homologous end joining 12,19.Despite these defects,Rag2c /c mice are not lymphoma-prone.
We reasoned that Rag2c /c p532/2double-mutant mice might dis-play genomic instability and lymphomagenesis,even in the context of intact classical non-homologous end joining.Consistent with previous reports 15,our Rag2c /c mice displayed partial developmental blocks in B and T lymphopoiesis because of a selective V-to-DJ rearrangement defect (Supplementary Fig.1).Rag2c /c animals,observed for up to 1year,showed no obvious signs of tumorigenesis (Fig.1a and data not shown).As expected 20,approximately two-thirds of p532/2mice developed thymic lymphoma at an average age of approximately 23weeks (mean survival 522.8weeks)(Fig.1a,b).Similar findings in RAG/p53-deficient mice 21demonstrate that RAG-initiated double-strand breaks are not critical initiators of lymphomagenesis in p53-deficient mice.In sharp contrast,100%(n 525)of our Rag2c /c p532/2mice died within 16weeks (mean survival 512.1weeks)with aggressive thymic lymphomas (Fig.1a–c).Tumour cells were highly proliferative and expressed cell surface CD4and CD8(Supplementary Fig.2),with little or no surface TCR (CD3e or TCR b )(data not shown),indicating that these tumours originate from immature thymocytes.
Tumours with highly proliferating lymphoblasts were detected in 4-to 6-week-old Rag2c /c p532/2thymi,but not in other organs (data not shown),confirming their thymic origin.Rag2c /c p532/2tumours generally displayed one or a few predominant D b 1–J b 1or D b 2–J b 2rearrangements,indicating a clonal or oligoclonal origin (Supplemen-tary Fig.3).
We next examined genomic stability in lymphomas from Rag2c /c p532/2mice,first by analysis of Giemsa-stained metaphase spreads prepared from 12Rag2c /c p532/2and two p532/2thymic lymphomas (Supplementary Table 1).Wild-type thymocytes showed almost no abnormal metaphases (0–3%)(Supplementary Table 1).In contrast,p532/2and Rag2c /c p532/2tumours harboured a variety of cytogenetic aberrations (aberrant metaphases:8–94%),including aneuploidy,chro-mosome breaks and chromosome fusions (Supplementary Table 1).We
1
Department of Pathology,New York University School of Medicine,New York,NY 10016,USA.2Department of Genetics,The M.D.Anderson Cancer Center,Houston,TX 77030,USA.3Department of Medicine,Division of Clinical Pharmacology,Center for Health Informatics and Bioinformatics,New York University School of Medicine,NY 10016,USA.4Department of Laboratory Medicine,Yale University School of Medicine,New Haven,CT 06520,USA.5Department of Immunology and Molecular Pathology,Division of Infection and Immunity,University College London,London W1T 4JF,
UK.
Time (weeks)
S u r v i v a l (%)Lymphoma Other tumour types
100%
67%
33%
Wild type
Rag2c/c ;p53–/–
1 cm
a
b
c
Rag2c/c ;p53–/–
(n = 25)p53–/–(n = 27)Figure 1|The C terminus of RAG2is a tumour suppressor in developing thymocytes.a ,Kaplan–Meier tumour-free survival analysis for cohorts of control (wild type,n 512;Rag2c /c ,n 519),p532/2(n 532)and Rag2c /c
p532/2(n 525)mice.Animals were monitored for 50weeks.The average age of death in weeks is shown for p532/2(22.8weeks)and Rag2c /c p532/2
(12.1weeks)genotypes with the P value determined by a Wilcoxon rank sum test.b ,Tumour spectrum observed for Rag2c /c p532/2(n 525)and p532/2mice (n 527).All Rag2c /c p532/2animals (n 525)showed enlarged thymus.p532/2animals showed either enlarged thymus and/or spleen (n 518)or other non-lymphoid tumour mass (n 59).c ,Physical appearance of normal thymus (wild type)and thymic lymphoma (Rag2c /c p532/2,arrow)of 3-month-old animals.
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analysed three Rag2c /c p532/2thymic lymphomas using spectral (1790T and 1745T)and G-band (1779T)karyotyping (Fig.2).We observed recurrent translocations involving chromosomes that harbour Tcr (chromosomes 14and 6)and Ig (chromosomes 12,6and 16)loci,suggesting that these might have been initiated by RAG-generated breaks.Moreover,all three lymphomas harboured translocations of the Igh locus-containing chromosome 12and/or the Tcr a /d locus-containing chromosome 14,loci that rearrange in thymocytes 22.Analysis of lymphoma 1779T revealed a C12;14translocation (Fig.2).These results suggest that Rag2c /c p532/2T cell tumours harbour clonal translocations involving the Tcr a /d and Igh loci,as seen in T-cell lym-phomas from patients with ataxia–telangiectasia and Atm 2/2mice 7,8,10,11,rearrangements not observed in p532/2lymphomas 21,23.To confirm the involvement of the Tcr a /d locus in chromosome translocations,we performed DNA fluorescence in situ hybridization (DNA FISH)analyses on metaphases from Rag2c /c p532/2thymic lymphomas (2489T and 2805T)using probes centromeric (Tcr a /d V )and telomeric (Tcr a /d C )to the Tcr a /d locus plus a paint for chro-mosome 14(Fig.3a).In both tumours,breakpoints within the Tcr a /d locus of one of the two chromosomes 14resulted in amplification of the Tcr a /d V region (Fig.3a).The telomeric fragment (including Tcr a /d C )was either translocated (2489T),or lost (2805T)(Fig.3a).DNA FISH analysis of tumours 1790T and 1779T (from Fig.2)using Tcr a /d C and V probes also confirmed translocation of chromosome 14with breakpoints within the Tcr a /d locus,although without obvious amp-lification (Supplementary Fig.4).
We next performed DNA FISH on Rag2c /c p532/2thymic lympho-mas 2489T and 2805T using probes centromeric (Igh C )and telomeric (Igh V )to the Igh locus along with a chromosome 12paint (Fig.3b).In both lymphomas,one chromosome 12showed translocation with another chromosome,with accompanying loss of both Igh C and V signals (Fig.3b).This could result from RAG-induced breaks with loss of the telomeric end of the chromosome (including Igh V )and loss of the Igh C region by end degradation before fusion to the partner chromosome,as previously reported in Atm 2/2mouse T cells 8.
Moreover,dual chromosome 12and 14paint analysis showed a C12;14translocation in lymphoma 2489T (Fig.3b).In contrast to Rag2c /c p532/2lymphomas,DNA FISH on metaphases from one p532/2thymic lymphoma (6960T)indicated that both Tcr a /d and Igh loci were intact (Supplementary Fig.5),consistent with previous work 21.
We next performed array-based comparative genomic hybridiza-tion (a-CGH)analysis on genomic DNA from five Rag2c /c p532/2thymic lymphomas (2489T,2805T,1348T,1779T,1780T).We observed loss or gain of a region within the Tcr a /d and Igh loci,reflect-ing V(D)J recombination (Supplementary Fig.6).All five Rag2c /c p532/2lymphomas examined showed substantial amplification of a common region on chromosome 14,centromeric of the Tcr a /d locus (Supplementary Fig.6a),in agreement with our FISH analyses (Fig.3a).We also observed loss of a common region on chromosome 12,telomeric of the Igh locus in all five Rag2c /c p532/2thymic lym-phomas analysed (Supplementary Fig.6b).Tumours 1779T,2489T and 2805also showed loss of a large region centromeric of the Igh locus,probably reflecting DNA-end degradation before fusion to the partner chromosome (Figs 2and 3a,b and Supplementary Fig.6b).In contrast,aCGH analysis of p532/2thymic lymphoma 6960T failed to reveal amplification centromeric to the Tcr a /d locus or deletion telo-meric to the Igh locus (Supplementary Fig.7a,b),in agreement with our FISH analysis (Supplementary Fig.5)and previous data 23.
Blocking lymphocyte development in early stages can lead to per-sistent RAG activity,which,in the absence of p53,can provoke lym-phomagenesis 23.To investigate whether the partial developmental block in Rag2c /c thymocytes 15is sufficient to produce genomic instab-ility and lymphomagenesis,we crossed core Rag1knock-in animals,which display diminished recombination and a strong block in B-and T-cell development 14,24(Supplementary Fig.1),into a p53-deficient background.Rag1c /c p532/2mice survived at an average age of 18.7weeks (Supplementary Fig.8a),barely distinguishable from p532/2mice.Also like p532/2mice,only two-thirds of Rag1c /c p532/2mice developed thymic lymphomas (Supplementary Fig.8b).
1745T
1790T
–X –6
–16
–12
–6
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–14
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Tumour number
Genotype Translocations*Frequency 1790
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t(9;12)7/8t(X;14)
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Rag2c/c ;p53–/–t(16;12)3/9t(6;6)
3/91779
Rag2c/c ;p53–/–t(12;14)
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Figure 2|Rag2c /c p532/2thymic lymphomas display recurrent
translocations involving chromosomes that harbour antigen-receptor loci.Representative images of spectral karyotyping (1790T and 1745T)and G-band karyotyping (1779T)analysis of three Rag2c /c p532/2T cell lymphomas.
Metaphase number analysed and translocations for each tumour sample are listed in the table.All three tumours harbour clonal translocations involving chromosomes that carry Tcr (chromosome 14,Tcr a /d ;chromosome 6,Tcr b )and/or Ig (chromosome 12,Igh ;chromosome 6,Ig k ;chromosome 16,Ig l )loci.
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Furthermore,metaphase DNA FISH analyses on two Rag1c /c p532/2thymic lymphomas (8383T and 8411T)(Supplementary Fig.9)and aCGH analysis on genomic DNA from four Rag1c /c p532/2thymic lymphomas (8315T,8333T,8383T,8411T)(Supplementary Fig.10)showed no evidence of recurrent translocations,genomic amplifica-tion or genomic deletion at chromosome 14and chromosome 12.The genomic instability observed in Rag2c /c p532/2thymic lymphomas is
therefore associated specifically with loss of the RAG2C terminus,and does not result from the developmental block in core RAG2homozygotes.We next asked whether core RAG2promotes genomic instability in the presence of p53by using three-dimensional interphase DNA FISH to examine the integrity of Tcr a /d locus (Fig.3c)in Rag2c /c double-positive thymocytes.The two alleles appeared as two pairs of signals (Tcr a /d V and Tcr a /d C ,mapping the two ends of the locus)in
most
Igh
Jh segments
Translocation
Chr. 12
Normal
Igh V Igh C
Translocation Normal Chr. 12Igh C
Chr. 12Chr. 14
Normal
Igh V Igh C
Translocation Normal Chr. 12Igh C Igh C
Igh V
b
E x a m p l e s o f R a g 2c /c n u c l e i w i t h a b e r r a n t T c r α/δ l o c i
Tcr α/δV
Tcr α/δC W i l d t y p e
c
WT
Rag2c/c
p53–/–
1234567N u m b e r o f a b e r r a n t c e l l s (%)
1.19 1.26.35
P < 0.001P < 0.001
d
Amplification Normal
Normal Tcr α C Tcr α V
Amplification Normal Normal Chr. 14Tcr α V
Amplification Translocation Normal
Tcr α C Tcr α V Amplification Translocation Normal Chr. 14Tcr α V Rag2c/c ; p53–/– (2489T)Rag2c/c ; p53–/– (2805T)Rag2c/c ; p53–/– (2489T)Rag2c/c ; p53–/– (2805T)Tcr α V
Tcr α C
V-D-J-V δTcr α/δ
a
Figure 3|Rag2c /c p532/2thymocytes display Tcr a /d -and Igh -associated genomic instability.a ,Top panel:schematic of the Tcr a /d locus,with positions of the BACs used for generation of DNA FISH probes indicated.Bottom panels:representative metaphases from two Rag2c /c p532/2thymic lymphomas using the Tcr a /d V BAC probe (red signal)combined with
chromosome 14paint (green signal,top row)or with the Tcr a /d C BAC probe (green signal,bottom row).Arrows point to the amplification of the Tcr a /d V region,arrowheads point to the translocated chromosome 14.b ,Top panel:schematic of the Igh locus,with positions of the BACs used for generation of DNA FISH probes indicated.Bottom panels:representative metaphases from the same two Rag2c /c p532/2thymic lymphomas using the Igh C BAC probe
(red signal)combined with chromosome 12paint (green signal,top row)or with the Igh V BAC probe (green signal,bottom row).Combination of
chromosome 12(red)and chromosome 14(green)paints is shown for both tumours in black boxes.Arrowheads point to the translocated chromosome 12.c ,Examples of confocal sections of three-dimensional Tcr a /d DNA FISH on freshly isolated wild-type (top row)or Rag2c /c (bottom rows)double-positive thymocytes.Tcr a /d V (green)and C (red)BAC probes were used.Scale bar,1m m.d ,Representative experiment showing the frequency at which Tcr a /d V and/or Tcr a /d C signals are lost in wild-type (WT),p532/2and Rag2c /c
thymocytes (n .200;see Supplementary Fig.11for additional experiments and statistical analysis).
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(.98%)wild-type and p532/2double-positive thymocytes (Fig.3d and Supplementary Fig.11),indicating that p53deficiency alone does not disrupt the integrity of the Tcr a /d locus,as expected 25.In contrast,Rag2c /c double-positive thymocytes displayed a three-to fivefold increase in the number of cells showing loss of at least one signal (Fig.3c,d and Supplementary Fig.11).These results suggest that core RAG2promotes genomic instability at the Tcr a /d locus,a phenotype similar to that previously reported in Atm 2/2and 53bp12/2animals 9,25.We noted that both Rag2c /c p532/2and Atm 2/2mice feature RAG-dependent genomic instability at the Tcr a /d and Igh loci,with develop-ment of pro-T cell lymphomas bearing clonal translocations,including 12/14translocations 2,3,7–11.To determine whether Atm 2/2thymic lymphomas also harbour amplification close to the Tcr a /d locus,we performed DNA FISH analysis for Tcr a /d and chromosome 14on metaphases from one Atm 2/2thymic lymphoma (10375T)(Sup-plementary Fig.12a).Both chromosomes 14showed translocations with breakpoints within the Tcr a /d locus,and amplification of the Tcr a /d V region on one allele (Supplementary Fig.12a),results that were confirmed by aCGH analysis (Supplementary Fig.12b).We also observed loss of DNA at a distal region of chromosome 12,near the Igh locus (Supplementary Fig.12b),as in Rag2c /c p532/2lymphomas (Fig.3a,b and Supplementary Fig.6).These data agree with recent analysis of thymic lymphomas from ATM-deficient mice 7.Thymic lymphomas that arise in other mutant backgrounds such as p53,core RAG1/p53(Supplementary Figs 8–10),E b /p53or H2AX/p53lack recurrent amplifications of chromosome 14regions and/or recurrent chromosome 12/14translocations,and thus appear to arise from dis-tinct mechanisms.
Our data reveal a novel in vivo function for the RAG2C terminus in promoting genomic stability.How does core RAG2allow genomic instability?We hypothesized that core RAG2,like the absence of ATM 3,destabilizes the post-cleavage complex.To investigate this,we generated RAG-signal end complexes by in vitro cleavage and challenged them at increasing temperatures,followed by gel electro-phoresis (Fig.4).Complexes containing full-length RAG2did not release 50%of signal ends until 55u C (Fig.4b,c),as expected 13,26.In contrast,core RAG2-containing complexes displayed statistically sig-nificant instability at lower temperatures,with 50%end release at 37u C (Fig.4b,c).To examine the post-cleavage complex in vivo ,we analysed
inversional recombination,which requires coordination of all four DNA ends.Decreased inversional recombination and increased formation of hybrid joints (generated by joining of a coding end to a signal end,in this case revealing defects in formation of four-ended inversion products)has been reported in ATM-and MRE11complex-deficient cells 3,27,28.As expected 3,28,we observed increased hybrid joint formation at the Ig k locus (V k 6-23to J k 1)in Atm 2/2and Nbs D B /D B splenocytes (Supplementary Fig.13).Importantly,we observed increased V k 6-23-to-J k 1hybrid joints in Rag2c /c splenocytes,com-pared with their wild-type and Rag2c /1counterparts (Supplemen-tary Fig.13).These results are supported by the observation that Rag2c /c lymphocytes exhibit defects in inversional recombination 15.Together,these data support our hypothesis that core RAG2impairs the stability of the RAG post-cleavage complex in vitro and in vivo .Our data support a common model for genomic instability in Rag2c /c p532/2and Atm 2/2mice:premature release of RAG-generated double-strand breaks from the RAG post-cleavage complex allows ends to escape the normal joining mechanisms,to persist and to be poten-tially joined by alternative non-homologous end joining,a pathway permissive for chromosome translocations and amplification 4,29.Both end release and end persistence are promoted by ATM deficiency 2,3,probably because ATM both stabilizes the RAG post-cleavage com-plex 3and activates p53-dependent checkpoints/apoptosis.In Rag2c /c p532/2mice,end persistence might be augmented by ongoing RAG activity through the cell cycle resulting from impaired degradation of core RAG2,which lacks the cell-cycle-regulated degradation motif 18,30.The complete penetrance,rapid development of lymphoma and extraordinary degree of RAG-mediated genomic instability make Rag2c /c p532/2mice an attractive model for investigating the spectrum of somatic genome rearrangements underlying lymphomagenesis.
METHODS SUMMARY
Mice.Mice were bred in the New York University Specific Pathogen Free facility;animal care was approved by the NYU SoM Animal Care and Use Committee (protocol number 090308-2).
Analysis of tumour cells.Lymphoid tumours were analysed by flow cytometry with antibodies against surface B-and T-cell markers.Metaphases were prepared and analysed as described in Methods.
FISH and image analysis.DNA FISH was performed using BAC probes as described in Methods.Interphase FISH was performed on
double-positive
a
b c
RAG1/RAG2
+ HMG + DNA substrate
37 °C
Signal end complex Challenge at increasing temperatures
Signal end release
Substrate
SC CE
SE
Full-length RAG2 Core RAG2
37°42°50°55°60°70°PK 37°42°50°55°60°70°PK
A v e r a g e e n d r e l e a s e (%)
Temperature (°C)
Figure 4|The C terminus of RAG2stabilizes the RAG post-cleavage
complex.a ,Biochemical end-release assay.Purified glutathione S -transferase (GST)-tagged core RAG1and non-tagged RAG2(full length or core)proteins (yellow circles)cleave a 500base pair (bp)DNA substrate at 37u C.Post-cleavage signal end complexes are thermally challenged at increasing temperatures to force the release of signal ends,which are detected after electrophoresis and gel staining.b ,Representative gel for end-release assays.
Numbers above each lane indicate the temperatures (in degrees Celsius)the reactions were heated to before electrophoresis.CE,coding ends;SC,single cleavages;PK,samples treated with proteinase K and SDS.c ,Quantification of signal end release,measured as the combined amount of signal ends divided by the signal from the total amount of DNA in the lane,from six experiments using two different protein preparations (*P ,0.05,Student’s t -test).
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thymocytes isolated by cell sorting according to protocols described in Methods. Images were obtained by confocal microscopy on a Leica SP5AOBS system,with optical sections separated by0.3m m.Images were analysed using Image J software. Metaphase spreads were imaged by fluorescent microscopy on a Zeiss Imager Z2 Metasystems METAFER3.8system and analysed using ISIS software.Statistical analysis of image parameters used a two-tailed Fisher’s exact test. Biochemical end-release assay.The stability of RAG-signal end complexes was measured as described in Methods.Briefly,RAG cleavage reactions were divided into aliquots in microfuge tubes and incubated at the indicated temperatures for 30min,followed by polyacrylamide gel electrophoresis.DNA was stained using SYBR Safe DNA Gel Stain(Invitrogen)and quantified with Quantity One software (Biorad).Student’s t-test assuming equal variance was used to calculate statistical significance.
aCGH analysis.For CGH,genomic DNA from mouse thymic lymphomas was profiled against matched thymic DNA from wild-type mice.aCGH experiments were performed on two-colour Agilent244A Mouse Genome Microarrays.Data analysis was performed as described in Methods.
Full Methods and any associated references are available in the online version of the paper at https://www.wendangku.net/doc/2a3943292.html,/nature.
Received6May;accepted15December2010.
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Supplementary Information is linked to the online version of the paper at
https://www.wendangku.net/doc/2a3943292.html,/nature.
Acknowledgements We thank M.Schlissel for the gift of core Rag2mice,F.Alt for the gift of core Rag1mice and S.Hewitt for the Igh BAC probes.D.B.R.was supported by National Institutes of Health Roadmap Initiative in Nanomedicine through a Nanomedicine Development Center award(1PN2EY018244),a National Institutes of Health grant CA104588and the Irene Diamond Fund.L.D.is a Fellow of The Leukemia and Lymphoma Society.A.V.A.was supported in part by grant1UL1RR029893from the National Center for Research Resources,National Institutes of Health.J.A.S.was supported by a National Institutes of Health grant R01GM086852,a National Institutes of Health Challenge grant NCI R01CA145746-01,a Leukemia and Lymphoma Scholar Award and a Wellcome trust project grant085096.
Author Contributions L.D.and D.B.R.conceived the study and co-wrote the manuscript.L.D.designed the experiments.L.D.,J.C.,M.C.and A.M.performed the experiments.Y.C.provided assistance with the mouse colonies.A.V.A.performed the aCGH data analysis.J.A.S.and S.C.provided technical and conceptual support.J.C.and J.A.S revised the manuscript.All the authors read and approved the manuscript. Author Information Reprints and permissions information is available at
https://www.wendangku.net/doc/2a3943292.html,/reprints.The authors declare no competing financial interests. Readers are welcome to comment on the online version of this article at
https://www.wendangku.net/doc/2a3943292.html,/nature.Correspondence and requests for materials should be addressed to D.B.R.(david.roth@https://www.wendangku.net/doc/2a3943292.html,).
LETTER RESEARCH
3M A R C H2011|V O L471|N A T U R E|123
METHODS
Mice.We obtained wild type(Taconic),Rag2c/c15,Rag1c/c24,p532/2(Jackson laboratory20)and Atm2/2(Jackson laboratory11)mice for this study.Rag2c/c or Rag1c/c mice were bred with p53-deficient mice to generate doubly deficient mice. Genotyping of these mutants was performed by PCR of tail DNA as described in the relevant references11,15,20,24.
Characterization of tumour cells and metaphase preparation.Lymphoid tumours were analysed by flow cytometry with antibodies against surface B-cell (CD43,B220,IgM)and T-cell(CD4,CD8,CD3,TCR-b)markers.FACS analysis used a BD LSRII flow cytometer(BD Biosciences)equipped with FacsDiVa and FlowJo.For metaphase preparation,tumour cells were prepared as previously described31,32.Briefly,primary tumour cells were grown in complete RPMI media for4h and exposed to colcemid(0.04m g ml21,GIBCO,KaryoMAX Colcemid Solution)for2hours at37u C.Then,cells were incubated in KCl75mM for 15min at37u C,fixed in fixative solution(75%methanol/25%acetic acid)and washed three times in the fixative.Cell suspension was dropped onto pre-chilled glass slides and air-dried for further analysis.
G-banding and spectral karyotyping.Optimally aged slides were treated for the induction of G-banding following the routine procedure33.Spectral karyotyping was performed using the mouse chromosome SKY probe Applied Spectral Imaging according to the manufacturer’s instructions to determine chromosomal rearrangements in the tumour samples.The slides were analysed using a Nikon Eclipse80i microscope.G-banding as well as SKY images were captured and karyotyped using an Applied Spectral Imaging system.
DNA FISH probes.BAC probes for the Igh and Tcr a/d loci were labelled by nick-translation and prepared as previously described34,35.For the Igh locus,BAC199 (Igh C)and BAC RP24-386J17(Igh V)were labelled in Alexa Fluor594and488 respectively(Molecular Probes).For the Tcr a/d locus,BAC RP23-304L21(Tcr a/d V)and RP-23255N13(Tcr a/d C)were labelled in Alexa Fluor488or594. StarFISH-concentrated mouse FITC or Cy3chromosome12or14paints were pre-pared following supplier’s instructions(Cambio).BAC probes were re-suspended in hybridization buffer(10%dextran sulphate,53Denharts solution,50%formamide) or in paint hyb buffer,denatured for5min at95u C and pre-annealed for45min at 37u C before hybridization on cells.
DNA FISH on metaphase spreads.Slides were dehydrated in ethanol series, denatured in70%formamide/23SSC(pH7–7.4)for1min30s at75u C,dehy-drated again in cold ethanol series,and hybridized with probes o/n at37u C in a humid chamber.Slides were then washed twice in50%formamide/23SSC and twice in23SSC for5min at37u C each.Finally,cells were mounted in ProLong Gold(Invitrogen)containing49,6-diamidino-2-phenylindole(DAPI)to counter-stain total DNA.
DNA FISH on interphase nuclei.Double-positive thymocytes were isolated from total thymi on a Beckman-Coulter MoFlo cell sorter as Thy1.21CD41CD81cells using the following antibodies:PE-Cy7-coupled anti-CD90.2(Thy1.2;53-2.1), APC-coupled anti-CD4(L3T4)and FITC-coupled anti-CD8(53-6.7).Cells were washed two times in13PBS and dropped onto poly-L-lysine-coated coverslips.For three-dimensional DNA FISH analyses,we used a protocol for immunofluor-escence/DNA FISH previously described34,35,with protein detection step omitted. Briefly,cells were fixed in2%paraformaldehyde/13PBS for10min at room temperature,permeabilized in0.4%Triton/13PBS for5min on ice,incubated with0.01mg ml21Rnase A for1h at37u C and permeabilized again in0.7%Triton /0.1M HCl for10min on ice.Cells were then denatured in1.9M HCl for30min at room temperature,rinsed in cold13PBS and hybridized overnight with probes at 37u C in a humid chamber.Cells were then rinsed in23SSC at37u C,23SCC at room temperature and13SSC at RT,30min each.Finally,cells were mounted in ProLong Gold(Invitrogen)containing DAPI to counterstain total DNA. Biochemical end-release assay.End-release assay to measure the stability of the signal-end complexes was performed as previously described26.For RAG-mediated cleavage,100ng of recombination substrate(PCR product from pJH289)was incubated for3h at37u C with200ng purified RAG protein and 200ng of purified recombinant HMGB1in a buffer containing50mM HEPES (pH8.0),25mM KCl,4mM NaCl,1mM DTT,0.1mg BSA,5mM CaCl2and 5mM MgCl2.Reactions were then divided into aliquots in microfuge tubes and incubated at different temperatures,or treated with stop buffer(10mM Tris(pH 8.0),10mM EDTA,0.2%SDS,0.35mg ml21proteinase K(Sigma Aldrich))for 30min and then run out on4–20%acrylamide tris-borate-EDTA(TBE)gels (Invitrogen).
aCGH analysis.aCGH experiments were performed on two-colour Agilent244A Mouse Genome Microarray.After internal Agilent quality control,the collected data were background subtracted and normalized using the Loess method36.We used circular binary segmentation method to define regions of copy number alteration compared with the control37and applied the cghMCR method for extraction of altered minimum common regions between the samples38.The analyses and visualizations were performed using the R statistical program39. 31.Theunissen,J.W.&Petrini,J.H.Methods for studying the cellular response to DNA
damage:influence of the Mre11complex on chromosome metabolism.Methods Enzymol.409,251–284(2006).
32.Multani,A.S.et al.Caspase-dependent apoptosis induced by telomere cleavage
and TRF2loss.Neoplasia2,339–345(2000).
33.Pathak,S.Chromosome banding techniques.J.Reprod.Med.17,25–28(1976).
34.Hewitt,S.L.et al.RAG-1and ATM coordinate monoallelic recombination and
nuclear positioning of immunoglobulin loci.Nature Immunol.10,655–664(2009).
35.Skok,J.A.et al.Reversible contraction by looping of the Tcra and Tcrb loci in
rearranging thymocytes.Nature Immunol.8,378–387(2007).
36.Yang,Y.H.et al.Normalization for cDNA microarray data:a robust composite
method addressing single and multiple slide systematic variation.Nucleic Acids Res.30,e15(2002).
37.Olshen,A.B.,Venkatraman,E.S.,Lucito,R.&Wigler,M.Circular binary
segmentation for the analysis of array-based DNA copy number data.Biostatistics 5,557–572(2004).
38.Aguirre,A.J.et al.High-resolution characterization of the pancreatic
adenocarcinoma genome.Proc.Natl https://www.wendangku.net/doc/2a3943292.html,A101,9067–9072(2004). 39.R.development Core Team.R:A Language and Environment for Statistical
Computing.Vienna:R Foundation for Statistical Computing(2006).
RESEARCH LETTER
对翻译中异化法与归化法的正确认识
对翻译中异化法与归化法的正确认识 班级：外语学院、075班 学号：074050143 姓名：张学美 摘要：运用异化与归化翻译方法,不仅是为了让读者了解作品的内容,也能让读者通过阅读译作,了解另一种全新的文化,因为进行文化交流才是翻译的根本任务。从文化的角度考虑,采用异化法与归化法,不仅能使译文更加完美,更能使不懂外语的人们通过阅读译文,了解另一种文化,促进各民族人们之间的交流与理解。翻译不仅是语言符号的转换,更是跨文化的交流。有时,从语言的角度所作出的译文可能远不及从文化的角度所作出的译文完美。本文从翻译策略的角度，分别从不同时期来说明人们对异化法与归化法的认识和运用。 关键词:文学翻译；翻译策略；异化；归化；辩证统一 一直以来,无论是在我国还是在西方,直译(literal translation)与意译(liberal translation)是两种在实践中运用最多,也是被讨论研究最多的方法。1995年,美籍意大利学者劳伦斯-韦努蒂(Lawrence Venuti)提出了归化(domestication)与异化(foreignization)之说,将有关直译与意译的争辩转向了对于归化与异化的思考。归化与异化之争是直译与意译之争的延伸,是两对不能等同的概念。直译和意译主要集中于语言层面,而异化和归化则突破语言的范畴,将视野扩展到语言、文化、思维、美学等更多更广阔的领域。 一、归化翻译法 Lawrwnce Venuti对归化的定义是,遵守译入语语言文化和当前的主流价值观,对原文采用保守的同化手段,使其迎合本土的典律,出版潮流和政治潮流。采用归化方法就是尽可能不去打扰读者,而让作者向读者靠拢(the translator leaves the reader in peace, as much as possible, and moves the author towards him)。归化翻译法的目的在于向读者传递原作的基本精神和语义内容,不在于语言形式或个别细节的一一再现。它的优点在于其流利通顺的语言易为读者所接受,译文不会对读者造成理解上的障碍,其缺点则是译作往往仅停留在内容、情节或主要精神意旨方面,而无法进入沉淀在语言内核的文化本质深处。 有时归化翻译法的采用也是出于一种不得已,翻译活动不是在真空中进行的,它受源语文化和译语文化两种不同文化语境的制约,还要考虑到两种文化之间的
高级英语写作黄金句型和新词
1) With the rapid improvement in.../growing awareness of..., more and more.../sth.... 1 随着…的飞速发展/越来越多的关注，越来越… (e.g. With the considerable improvement in building industry, more and more structures are being erected to set the people's minds at ease.) 例：随着建筑业的大力推进，人们对建立越来越多的房屋建筑感到宽心。 2) Recently, sth./the problem of...has been brought to popular attention/ has become the focus of public concern. 2 近来，某事/某问题引起了人们的普遍关注/成了公众关注的焦点。 (e.g. Recently, the problem of unemployment has been brought to such popular attention that governments at all levels place it on the agenda as the first matter.) 例：近来失业问题引起了人们的普遍关注，各级政府已把它列为首要议程。 3) One of the universal issues we are faced with/that cause increasing concern is that... 3 我们面临的一个普遍问题是… /一个越来越引人关注的普遍问题 是… (e.g. One of the universal issues that draw (cause) growing concern is whether it is wise of man to have invented the automobile.) 例：一个越来越引人关注的普遍问题是，发明汽车是否为人 4) In the past few years, there has been a boom/sharp growth/decline in.. . 4 在过去的几年里，…经历了突飞猛进/迅猛增长/下降。 (e.g. In the past ten years, there has been a sharp decline in the number of species.) 例：在过去的几年里，物种数量骤然下降。 5) Nowadays, more/most important/dangerous for our society is... 5 如今对我们社会更（最）为重要的（危险的）事情是… (e.g. Nowadays, most dangerous for our society is the tendency to take advantage of each other in political circles.) 例：对我们社会最为危险的事情是政界倾向于互相利用。 6) According to the information given in the table/graph, we can find that... 6 根据图表资料，我们可以发现… 7) As can be seen from the table/graph/figure, there is a marked increase /decline/favorable (an unfavorable) change in... 7 根据图表（数字）显示，…明显增长（下降）/发生了有利（不利）变化。 8) As we can see from the table/graph/figure above, drastic/considerable/ great changes have taken place in...over the period of time from...(年份)to...( 年份) 8 据上面图表（数字）所示，从某年到某年某方面发生了剧烈的（相当大的；巨大的）变化。
Tube, pipe, tubing 和 piping 四个名词(术语)的区别
Tube, pipe, tubing 和 piping 四个名词（术语）的区别 CACI 总部 曹良知 ASME 锅炉及压力容器规范对tube, pipe, tubing 和piping 这四个名词（术语）的含义是很严密的，适用于不同的场合。现将我对四个名词的理解分述如下，供大家讨论。 1、 tube 是圆形的，或具有连续周边的任何其他截面形状的空心制品。圆形 tube 的尺寸 可以用外径、内径、壁厚三者中的任意两个来指定。 2、 pipe 是符合ASNI B36.10和B36.19（用于不锈钢）所列公称尺寸的圆形截面的tube , 它的直径用NPS 号表示，NPS 号与实际外径是不一致的，如NPS8 其外径是8.625 英寸。管子壁厚用schedule No. 表示，同一NPS 号可以有各种Sch. No.。Sch. No. 有标准壁厚（STD ）、加厚壁厚（XS ）和特厚壁厚（XXS ）之分等等。 因此，tube 和 pipe 的基本差别是制造所依据的尺寸标准。其次pipe 只有圆管，tube 可以有各种截面形状，pipe 只是tube 中的一个特例。第三，无论tube 还是pipe 都是指材料生产厂生产出的原材料。 3、 tubing 是用tube 按PG-27.2.1[外径D ≤5in.(127mm)]的计算公式计算后选定直径和 厚度，并按设计要求加工（如弯管、开坡口、甚至油漆等）后制成的产品。我们可以叫它管子件。 其计算公式是： e D P S PD t +++=05.02 ])005.0(201.02[e D t D e D t S P -----= 4、 piping 一般选用pipe 按PG-27.2.2的计算公式计算后选定NPS 和Sch. No ，并按设计 要求加工后制成的产品，我们称作管道，但锅炉制造厂一般将这些管子叫做导管。 其计算公式是： PD t = 或 c PR t += )(2)(2c t y D c t SE P ---= 或 ))(1() (c t y R c t SE P --+-= tubing 和piping 的区别是 1.选用的原材料不同，tubing 一般选用tube 加工制造piping 选用pipe 加工制造。2.计算用的公式不同。在这里我们不讨论计算公式的差别。而tube, pipe 和 tubing, piping 的区别。前者是生产厂的原材料，而后者是锅炉制造厂或压力容器制造厂制成的产品。这就是锅炉及压力容器规范的规则对这四个名词（术语）的含义和用法加以区分的要点。当然输送流体并已安装就位的管子也叫管道。所以我们对这四个名词的译名应区别开，以免混淆造成理解上的错误。特别是对外国人（如授权检验师），若把管子件（tubing ）说成管子（tube ），他就会疑惑不解。所以，建议把tube 译为管子，pipe 译成公称管；tubing 译成管子件，piping 译成管道（最好将工厂制成的产品译成导管，已安装好的叫管道，以便区别）。 以上是我个人的意见，若有不妥之处，请大家批评指正。
景区讲解员工作总结
景区讲解员工作总结 景区讲解员工作总结一 XX年是不平凡的一年，XX年我从学校走了出来，把两年里所学到的关于导游 的知识运用到我的工作中，从理论转向实践。XX年6月开始我在南岳衡山从事 地接导游导游工作，时间不长，资力也不深，而感慨却颇多：“导游”工作给我的 生活带来了许多快乐，却也让我知道，“导游”不是一项简单的工作，与其他职业 有一个显著的不同，那就是你必须与客人近距离接触，这自然使我们对服务的感 触比一般人深刻。从某种意义上可以这么讲，导游职业的无穷魅力正是源于我们 对服务的感知和热爱。 通过几个月的工作实践，我深深的体会到，取得了导游证，并不代表你就永远 是一个合格的导游员，而是要不断的的学习、充实、提高。在旅游者的眼中，导 游员应该是无所不知的“万事通”。导游服务是知识密集型的高智能的服务工作， 丰富的知识、广博的见闻是做好导游服务工的前提。作为一个导游员就要“与时俱进”，永远保持积极的求知欲，以适应社会进步和发展的需求。更重要的是。我们 自己千万不敢把自己当成“万事通”，要保持谦虚谨慎的态度，要切记“学海无涯”、“学无止境”，“人外有人，天外有天”，“三人行，必有我师”。 要时刻牢记导游的职责，认真学习《导游人员管理暂行规定》、《中华人民共 和国国家标准导游服务质量》，努力的实施好旅游计划，作好联系、协调、讲解 等服务工作。坚持“宾客至上、服务至上、为大家服务、合理而可能”的四大服务 原则细致、热心、周到的作好导游服务工作。也就是一切工作以旅游者为出发点，以服务为出发点，时刻考虑旅游者的利益和要求，绝不能拒绝游客的合理合法要求。服务过程中要坚持“为大家服务”的原则，不能有亲疏之分，厚此薄彼，而应 对每个游客都热情、周到、友好、尊重，不偏不倚、一视同仁;要坚持“合理而可能”的原则，在旅游服务过程中，要时刻关注游客的情绪变化，耐心倾听旅游者的 意见、要求，冷静分析、仔细甄别，合理又能实现的，就努力的去做，如果没有 作好或是已经错过机会，就想办法及时弥补，以求最大限度的达到游客的满意。 导游讲解服务是整个旅游服务活动过程中极为重要的一个方面，在导游讲解过程中，我认为“准确、清楚、生动”三者相辅相成，缺一不可，首先“准确”是首当其冲，至关重要的，在讲解过程中牢记“一伪灭千真”的教训，切忌胡编乱造、张冠 李戴、信口开河，这样会使游客有被蒙蔽、愚弄的感觉，会引起游客的反感、责备。旅游者在旅游活动中“求知”是重要的内容之一，而我们导游就起着传播知识 信息、传递审美观念、播洒中华文明的重任，因此导游语言必须科学、规范，传 递的信息必须正确无误，这样更能够吸引游客的注意，满足游客的“求知”愿望。 其次，“清楚”是关键，在导游讲解中，清楚、简洁流利的语言表达，是导游语言 科学性的又一体现。口齿清楚、言简意赅、措词恰当、组合相宜、层次分明、逻
我国最新无缝钢管行业术语及中英文对照
我国最新无缝钢管行业术语及中英文对照 eamless steel pipe 无缝钢管 Hot-finished steel pipe热轧钢管 Stainless steel pipe不锈钢钢管 Pipe fittings管件 对焊管件butt-welded pipe fitting （弯头elbow、等径和异径三通equal and reducing tees、同心和异心大小头concentric and eccentric reducer、 Flange 法兰（对焊法兰、平焊法兰、 Supporting tubing 管道配管 Hot-rolling seamless steel tube 热轧无缝钢管 Cold-drawn seamless steel tubes 冷拔无缝钢管 spiral steel pipe 螺旋钢管 Standard标准welded 焊接 Billet 管坯 Outside diameter 外径Outside diameter tolerance 外径差 Outside diameter thickness 外径厚度Wall Thickness Tolerance 壁厚差 Grade 钢级 Tensile strength 抗拉强度 Yield strength 屈服强度 Elongation 伸长 Impact 冲力 Hardness 硬度 Wall thickness 壁厚 Inspection 检验验收picking 酸洗grinding 磨修cooling 冷却heating 加热 生产设备production equipment 检测设备detection equipment x-ray detection 射线探伤 ultrasonic inspection 超声波探伤 hydrostatic testing fluid pipe 流体管 drill pipe 钻管drill rig 钻孔机hydraulic pipe 液压管 boiler pipe 高压炉管gas pipe 天燃气管 oil pipe石油管structure pipe 结构管 metal cutlery 金属餐具gas barbecue stove 燃气烧烤炉 无缝钢管Seamless Steel Pipe 锅炉管Boiler Tube 石油套管Pipe For Oil Field 高压管Pressure Pipe 高压气瓶用管Tube for High-Pressure Vessel 地质钻探用管Seamless Steel Pipe For Geological Drill In 本文由钢管世界-无缝钢管网提供：https://www.wendangku.net/doc/2a3943292.html,/转载注明出处!
高级英语一
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管件常用英文词汇及缩写I
弯头joint 弯头angle fittings; bent pipe; elbow; knee 弯头扳手bent-handle wrench; bent spanner 弯头车刀angular tool 弯头持针钳curved needle holders 弯头刀架knee tool 弯头道钉brob 弯头管swan neck 弯头键gib head key 弯头接合knee-joint 弯头螺栓bolt with one end bent back 弯头锁紧螺母elbow jam unt 弯头套管elbow union 弯头套筒扳手offset socket wrench 弯头脱水器elbow separator 弯头芯盒elbow core box 活接 活接loose joint 活接地swinging earth 活接结合articulated joint 活接联接器joint coupling 活接三通union tee 活接头articulation; union swivel; union 活接头螺母union nut 活接头螺栓swing bolt 活接头套管union thimble 活接卸槽articulated chute (浇混凝土用的) 三通 三通tee bend; tee branch; tee joint; tee conneetion; tee fitting; wye; Y; yoke 三通玻璃栓three-way glass stopcock 三通道放大器three-channel amplifier; three-path amplifier 三通道自动驾驶仪three-dimensional autopilot 三通电缆分线盒three-way coupling box 三通阀tee valve; three-port valve; three-way valve; triple valve 三通阀盖triple-valve cap 三通阀活塞triple-valve piston 三通阀联箱three way valve menifold
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八、讲解员讲解费由售票处统一收取，月底结算，严禁讲解员擅自违规收费。 九、讲解员在服务过程中要主动做好地貌保护的宣传和现场维护工作。 十、在讲解结束后应送游客到景区门口，并致欢送词。 十一、讲解员不得与游客发生争执，遇到特殊情况沉着应对并及时向领导反映，避免与游客发生冲突。 十二、讲解员违反本管理办法的，将给予每次20元—100元罚款；脱岗1次，罚款20元；因讲解员失误发生投诉案件的，每次处100元罚款；同时责令其限期改正，对逾期不改正或情节严重的，建议调整岗位。 十三、本办法由景区运营部负责解释。 十四、本办法自颁布之日起执行。
翻译中的归化与异化
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高级英语写作1-10课翻译
高级英语写作1-10课翻译
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景区讲解员的服务流程
景区讲解员的服务流程-标准化文件发布号：（9456-EUATWK-MWUB-WUNN-INNUL-DDQTY-KII
景区讲解员的服务流程 1准备工作 熟悉接待计划：旅游团的基本信息，旅游团成员的情况，交通工具，是否有特殊要求和注意事项。 落实接待事宜：落实旅游车辆、住宿及用餐，掌握全陪或者司机的联系电话。与他们联系商定第二天的接团时间及地点。 物质准备：游客接待确认书，游客意见单，话筒，导游旗。 形象准备：上团前要做好仪容、仪表方面的准备。整洁、大方、自然、不浓妆艳抹。 语言和知识准备：要在大脑里面准备好第二天要讲解的内容；接待有专业知识的团队，要做好相关的专业知识、词汇的准备工作。语言要生动、流畅和清楚。 心理准备：我们要能有技巧的回答客人提出的任何问题，面对客人的指责、抱怨，我们要冷静、沉着的面对。 联络畅通准备：上团前一天，要把手机，话筒的电充足，随时保持畅通。 2接团服务 出发前的准备：在旅行团到达之前，打电话给全陪或者司机，前一天晚上20：00前确定好到达时间、人数、停车位置、集合地点等，在集合地点恭候旅游团的到来。旅游团到达后，协助全陪或者司机购票，提醒客人山顶温度较低，指引乘客上车，清点一下车上的人数。检票后通知司机师傅开车。 途中导游：致欢迎词，山路崎岖，提醒客人把车辆扶手打下并系好安全带，当日活动安排（参观景点名称、途中所经地点、所需时间），介绍游览景点（讲安全讲游客接待中心，讲内循环通道，讲景区公交车的发班时间，讲游览线路，讲中途会经过哪几个景点，讲三条游步道），活跃气氛（做些娱乐互动，与游客互动） 景点讲解：抵达景点下车之前，告知游客车牌号、停车地点、开车时间、游览线路、游览所需时间、游览过程中的注意事项（不能随便雕刻、不能在景区乱扔垃圾、不能在景区内吸烟、不能随意摘取野果实，以免误食）等。抵达景点后，组织客人有顺序的下车，提醒客人注意脚下的安全，带领游客沿着游览线路对所见景物进行精彩的导游讲解。在游览过程中要随时随地注意游客的安全，特别是老弱病残的游客，要防止游客走失和意外事件的发生。 3送团服务 清点团队人数，后提醒客人清点一下随身携带物品，如无遗漏则请司机开车离开。致欢送词（感谢语、惜别语、征求意见语、致歉语、祝愿语）。致完欢送词后，将“游客意见反馈表”发给游客，请其填写。游客填写完毕后如数收回，妥善保留。 4总结工作 认真做好带团小结，实事求是的汇报接团情况。 2
翻译的归化与异化
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翻译的归化与异化 作者：熊启煦 作者单位：西南民族大学,四川,成都,610041 刊名： 西南民族大学学报（人文社科版） 英文刊名：JOURNAL OF SOUTHWEST UNIVERSITY FOR NATIONALITIES（HUMANITIES AND SOCIAL SCIENCE） 年，卷(期)：2005,26(8) 被引用次数：14次 参考文献(3条) 1.鲁迅且介亭杂文二集·题未定草 2.刘英凯归化--翻译的歧路 3.钱钟书林纾的翻译 引证文献(15条) 1.郭锋一小议英语翻译当中的信达雅[期刊论文]-青春岁月 2011(4) 2.许丽红论汉英语言中的文化差异与翻译策略[期刊论文]-考试周刊 2010(7) 3.王笑东浅谈汉英语言中的差异与翻译方法[期刊论文]-中国校外教育（理论） 2010(6) 4.王宁中西语言中的文化差异与翻译[期刊论文]-中国科技纵横 2010(12) 5.鲍勤.陈利平英语隐喻类型及翻译策略[期刊论文]-云南农业大学学报(社会科学版) 2010(2) 6.罗琴.宋海林浅谈汉英语言中的文化差异及翻译策略[期刊论文]-内江师范学院学报 2010(z2) 7.白蓝跨文化视野下文学作品的英译策略[期刊论文]-湖南社会科学 2009(5) 8.王梦颖探析汉英语言中的文化差异与翻译策略[期刊论文]-中国校外教育（理论） 2009(8) 9.常晖英汉成语跨文化翻译策略[期刊论文]-河北理工大学学报（社会科学版） 2009(1) 10.常晖对翻译文化建构的几点思考[期刊论文]-牡丹江师范学院学报（哲学社会科学版） 2009(4) 11.常晖认知——功能视角下隐喻的汉译策略[期刊论文]-外语与外语教学 2008(11) 12.赵勇刚汉英语言中的文化差异与翻译策略[期刊论文]-时代文学 2008(6) 13.常晖.胡渝镛从文化角度看文学作品的翻译[期刊论文]-重庆工学院学报(社会科学版) 2008(7) 14.曾凤英从文化认知的视角谈英语隐喻的翻译[期刊论文]-各界 2007(6) 15.罗琴.宋海林浅谈汉英语言中的文化差异及翻译策略[期刊论文]-内江师范学院学报 2010(z2) 本文链接：https://www.wendangku.net/doc/2a3943292.html,/Periodical_xnmzxyxb-zxshkxb200508090.aspx
英语专业(高级翻译)简介
英语专业(高级翻译)简介 ·日期：2007-06-07 ·点击次数： 1446 培养目标：本专业培养具有扎实的英语语言基础，较强的语言交际能力和跨文化交际能力，掌握多方面的翻译知识和技巧，能熟练地运用英、汉语在外事、外交、经贸、文化、科技等部门从事口译、笔译的专门人才。 知识能力 要求：1．语音、语调准确，清楚自然。 2．词法、句法、章法(包括遣词造句与谋篇布局)规范、表达得体。 3．认知词汇10000~12000，且能正确而熟练地使用其中的5000~6000个单词及其最常用的搭配。 4．听、说、读、写、译技能熟练，具有较强的英语综合运用能力。毕业时达到英语专业八级水平。 听的能力：听懂真实交际场合中各种英语会话；听懂英 语国家广播电台以及电视台(如VOA, BBC, CNN)有关政治、经济、文化、教育、科技 等方面的新闻、现场报道、专题报道、综合 评述以及与此类题材相关的演讲和演讲后 的问答；听懂电视时事报道和电视短剧中的 对话。语速为每分钟150~180个单词，理解 准确率不低于60%。 说的能力：能就国内外重大问题与外宾进行流利而得体 的交流；能系统、深入、连贯地发表自己的 见解。 读的能力：能读懂一般英美报刊杂志上的社论和书评、 英语国家出版的有一定难度的历史传记和 文学作品；能分析上述题材文章的思想观 点、语篇结构、语言特点和修辞手法。能在
5分钟内速读1600词左右的文章，掌握文章 的主旨和大意，理解事实和细节。 写的能力：能写各类体裁的文章，做到内容充实，语言 通顺，用词恰当，表达得体。写作速度为30 分钟300~400个单词。能撰写长度为 5000~6000个单词的毕业论文，要求思路清 晰、内容充实、语言通顺。 译的能力：1)笔译：能运用翻译的理论和技巧，将英美报 刊上的文章、文学、科技、文化、经贸方面 的原文译成汉语，或将我国报刊、杂志上的 文章、一般文学作品、科技、文化、经贸方 面的原文译成英语，速度为每小时250~300 个单词。译文要求忠实原意，语言流畅。 2)口译：掌握连续或同步口译所需的技巧及基 本要求，能担任外经外贸、外事、外交、国 际文化、科技交流活动的口译和国际会议的 同声传译任务。 5．具有宽广的英语专业知识和相关学科知识。英语专业知识包括文学、语言学和对象国社会与文化的知识。相关学科的知识涉及教育学、教育心理学、语言习得理论、英语教学法、英语测试理论与实践、课程设置等领域。6．具有获取知识的能力、运用知识的能力、分析问题的能力、独立提出见解的能力和创新的能力。 7．熟悉中、英两种文字计算机基本技术并能实际应用。 主干课程：基础英语、高级英语、散文选读、英语泛读、英语口语、基础英语写作、高级英语写作、英汉/汉英口译、英汉笔译、英语视听说、英语报刊选读、英美文学，同声传译、连续传译、经贸法律翻译等。 修业年4年
旅游景区讲解员讲解服务标准
旅游景区讲解员讲解服务标准 1范围 本标准规定了XX风景名胜区随车讲解员服务应达到的目的和要求。 本标准适用于XX风景名胜区随车讲解员讲解工作。 2 服务目的 为游客提供随车讲解服务，方便游客游览。 3 服务要求 3.1 时效性讲解人员应提前10分钟到岗，做好各项准备工作。 3.2 准确性 3.2.1 车辆出发前简单告知游客乘车注意事项，待游客坐稳后告知司机发车。 3.2.2 讲解员在讲解过程中要详细讲解景区的总体概况、沿途风光及景区各服务区基本情况，不得擅自减少或变更讲解内容。 3.2.3 在游客乘车过程中，告知游客景区投诉电话、游览注意事项等信息，当车辆快到站时，应提前准确报站。 3.3 文明性 3.3.1 在车辆启动的同时，致欢迎词：“尊敬的游客朋友，你们好，欢迎您来到XX世界地质公园观光游览”，介绍本人和司机师傅，公布景区投诉电话号码，表示诚挚服务的愿望，告知门票及安全注意事项。
3.3.2 讲解员举止大方得体，讲解内容生动形象，在沿途讲解中，不能随意减少讲解内容，严禁掺杂庸俗下流的内容，更不能接听电话，与人闲谈打闹。 3.3.3 讲解员处理各种事情要以大局为重，时刻维护景区利益与游客合法权益。耐心细致地解答游客提出的问题，想游客所想，急游客所急，主动为游客排忧解难。 3.4 着装要求讲解员须统一着装、佩戴工牌，保持良好的仪容仪表，穿着朴素大方，严禁穿奇装异服、浓妆艳抹，任何时候不得在游客面前整理衣裤。 3.5 行为要求讲解时要面对游客，站立服务，表情要自然、诚恳、和蔼，语言准确、生动、富有表达力，同时注意使用礼貌用语。严禁掺杂庸俗下流的内容，不得私自接听或拨打与工作无关的电话。 3.6 功能性告知游客使用门票的注意事项，建议游客合理安排旅游线路，介绍沿途风光及各服务区状况，提前报站，下车时提醒游客携带好随身物品。
归化与异化翻译实例
翻译作业10 Nov 15 一、请按归化法(Domestication)翻译下列习语。 Kill two birds with one stone a wolf in sheep’s clothing strike while the iron is hot. go through fire and water add fuel to the flames / pour oil on the flames spring up like mushrooms every dog has his day keep one’s head above water live a dog’s life as poor as a church mouse a lucky dog an ass in a lion’s skin a wolf in sheep’s clothing Love me, love my dog. a lion in the way lick one’s boots as timid as a hare at a stone’s throw as stupid as a goose wet like a drown rat as dumb as an oyster lead a dog’s life talk horse One boy is a boy, two boys half a boy, and three boys nobody. Man proposes, God disposes. Cry up wine and sell vinegar (cry up, to praise; extol: to cry up one's profession) Once bitten, twice shy. An hour in the morning is worth two in the evening． New booms sweep clean. take French leave seek a hare in a hen’s nest have an old head on young shoulder Justice has long arms You can’t teach an old dog Rome was not built in a day. He that lives with cripples learns to limp. Everybody’s business is nobody’s business. The more you get, the more you want. 二、请按异化法(foreignization)翻译下列习语。 Kill two birds with one stone a wolf in sheep’s clothing