日本血吸虫单克隆抗独特型抗体NP30轻链可变区基因的克隆及序列分析  

CLONING AND SEQUENCE ANALYSIS OF THE LIGHT CHAIN VARIABLE REGION GENE OF MONOCLONAL ANTI-IDIOTYPIC ANTIBODY NP30 OF SCHISTOSOMA JAPONICUM

作　　者：宋晓彤[1] 冯振卿[1] 仇镇宁[1] 李芸茜[1] 俞小淙[2] 熊英[2] 尹长城[2] 黄华梁[2] 管晓虹[1] 

机构地区：[1]南京医科大学医学分子生物研究所,南京210029 [2]中国科学院遗传研究所,北京100101

出　　处：《中国寄生虫学与寄生虫病杂志》2000年第5期257-259,共3页Chinese Journal of Parasitology and Parasitic Diseases

基　　金：国家自然科学基金!资助项目 (39970 6 70 )&&

摘　　要：[目的 ]分离日本血吸虫单克隆抗独特型抗体NP30轻链可变区 (VL)基因并测定其序列。 [方法 ]根据鼠免疫球蛋白轻链可变区基因FR1和FR4序列的保守性 ,化学合成用于体外扩增Ig轻链可变区基因的数对引物。以日本血吸虫单克隆抗独特型抗体NP30的杂交瘤细胞株基因组DNA为模板 ,扩增VL 基因 ,将其克隆入 pUC19载体 ,重组子用Sanger的双脱氧链终止法测定序列 ,将序列与GenBank中已发表的抗体序列比较。 [结果 ]VL 基因全长 318bp ,属鼠免疫球蛋白κ轻链第IV亚类 ,由种系基因V与Jκ4 重排而来。该VL 基因序列已被GenBank收录 (accessionNo AF2 0 6 72 0 )。 [结论 ]该VL 基因为日本血吸虫单克隆抗独特型抗体NP30轻链可变区基因。Objective] To amplify and sequence the light chain of anti idiotypic monoclonal antibody NP30 of Schistosoma japonicum. [Methods] By comparing the conserved regions at each end of the nucleotide sequences of murine germ line genes enco ding FR1 and FR4 regions of immunoglobulin light chain variable regions, we designed a set of primers for amplification of V L gene. The hybridoma cells secreting anti idiotypic monoclonal antibody NP30 of Schistosoma japonicum were cultured and their genome DNAs were extracted and used as templates for PCR. The PCR product was then cloned into pUC19 vector. The recombinants were sequenced by Sanger′s method. The V L gene was compared with GenBank and published mouse V L genes. [Results] The full length of V L gene was 318 bp. The V L gene was a member of mouse Ig κ light chain subgroup IV and generated from rearrangement of germ line V and Jκ 4 genes. The V L gene sequence has been registered by GenBank(accession No. AF206720). [Conclusion] The obtained V L gene was a potentially functional gene of anti idiotypic monoclonal antibody NP30 of Schistosoma japonicum .

关 键 词：日本血吸虫 抗独特型抗体 单克隆抗体 基因克隆 

分 类 号：R383.24[医药卫生—医学寄生虫学]