中国寄生虫学与寄生虫病杂志 ›› 2000, Vol. 18 ›› Issue (5): 1-259.

• 论著 •    下一篇

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

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

  

  1. 南京医科大学医学分子生物研究所,南京 210029 中国科学院遗传研究所,北京 100101
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2000-10-30 发布日期:2000-10-30

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

SONG Xiao tong 1;FENG Zhen qing 1;QIU Zhen ning 1;LI Yun qian 1;YU Xiao cong 2;XIONG Ying 2;YIN Chang cheng 2;HUANG Hua liang 2;GUAN Xiao hong 1
  

  1. 1 Institute for Molecular Biology of Medicine,Nanjing Medical University,Nanjing 210029 2 Institute of Genetics,Chinese Academy of Sciences,Beijing 100101
  • Received:1900-01-01 Revised:1900-01-01 Online:2000-10-30 Published:2000-10-30

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。

关键词: 日本血吸虫, 抗独特型抗体, 单克隆抗体, 基因扩增, 序列分析

Abstract:  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 .

Key words: Schistosoma japonicum, anti idiotypic antibody, gene amplification, sequence analysis