Cloning and sequence analysis of V_H and V_L gene of anti-CD3 McAb
- VernacularTitle:抗CD3单克隆抗体重、轻链可变区基因的克隆及序列分析
- Author:
Nan ZHANG
;
Mingjie CHEN
;
Jing CHEN
;
Al ET
- Publication Type:Journal Article
- Keywords:
CD3 McAb WuT3 Gene cloning DNA sequence analysis
- From:
Chinese Journal of Immunology
2000;0(09):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To amplify and sequence of the variable region genes of anti CD3 McAb.Methods:The V H?V L genes were amplified by RT PCR from total RNA that were extracted from WuT3 hybridoma.Recombinant cloning vector was constructed and sequenced after the enzyme digestion.Results:It showed that V H gene consisted of 363 bp encoding amine acid residues,belongs to mouse heavy chain subgroup IIB;V L gene consisted of 330 bp encoding amine acid residues,belongs to mouse ? light chain subgroup III.Comparing with Kabat database,the V H?V L genes were in agreement with the characterization of DNA sequences present in the mouse Ig V H?V L regions respectively.Conclusion:The success of cloning of the V H?V L genes of WuT3 McAb lay a good foundation for the construction and expression of chimeric antibody.
