摘要
目的 :抗CD3单克隆抗体 (WuT3)可变区基因克隆及序列分析。方法 :采用RT PCR技术 ,从WuT3杂交瘤细胞总RNA中扩增VH 、VL 片段 ,经酶切后通过链接反应构建重组克隆载体 ,测序鉴定。结果 :通过国际联机检索发现VH 、VL 基因与Ig同源 ,分别符合小鼠IgVH 、Vκ基因特征。VH 基因属于鼠重链VH 第IIB亚类 ,全长 36 3bp ,可编码 12 1个氨基酸 ;VL 基因属于鼠κ轻链第III亚类 ,全长 330bp ,可编码 110个氨基酸。结论 :成功获得WuT3单抗的重、轻链可变区基因。
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 κ 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.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2002年第9期608-610,共3页
Chinese Journal of Immunology
