Sequence and analysis of human anti-HBsκ light chain gene

SHI Xiaofeng; et al

doi:10.13406/j.cnki.cyxb.2004.05.010

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Sequence and analysis of human anti-HBsκ light chain gene

更新时间：2024-01-17

- Sequence and analysis of human anti-HBsκ light chain gene
- Journal of Chongqing Medical University Issue 5, Pages: 594-596(2004)
- 作者机构：
  
  1. 重庆医科大学病毒性肝炎研究所
  2. 重庆医科大学病毒性肝炎研究所,重庆,400010
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.2004.05.010
  CLC： R346
- Published：2004，
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[1]石小枫,刘杞,王志毅.人抗HBs轻链基因的序列测定及序列分析[J].重庆医科大学学报,2004(05):594-596+650.

SHI Xiaofeng, et al. Sequence and analysis of human anti-HBsκ light chain gene. [J]. Journal of Chongqing Medical University (5):594-596(2004)
[1]石小枫,刘杞,王志毅.人抗HBs轻链基因的序列测定及序列分析[J].重庆医科大学学报,2004(05):594-596+650. DOI： 10.13406/j.cnki.cyxb.2004.05.010.

SHI Xiaofeng, et al. Sequence and analysis of human anti-HBsκ light chain gene. [J]. Journal of Chongqing Medical University (5):594-596(2004) DOI： 10.13406/j.cnki.cyxb.2004.05.010.

摘要

目的 :获得人源性乙型肝炎病毒抗HBs轻链基因。方法 :从已构建的人源性噬菌体抗体库中

用固相化HBsAg对构建的人源性噬菌体抗体库进行三轮淘筛

经ELISA实验

筛选到抗HBs噬菌体抗体（P/N :2 .4 15 ）的阳性克隆

根据已知序列合成一对轻链引物

进行PCR扩增

扩增出轻链目的基因片段

将目的基因片段定向克隆入质粒pUC18

进行序列测定及分析。结果 :筛选出的克隆具有HBsAg特异性亲和力

构建了pUCl8-抗HBsκ链重组质粒

序列分析其为κ链

为 6 4 3bp包括了可变区和恒定区。结论 :利用抗原抗体特异性反应从噬菌体抗体库中筛选到抗HBs轻链基因。

Abstract

Objective:To obtain the clone and gene of light chain on human antibody against HBs.Methods:The phage displaying library was subjected to three rounds panning of “adherence-elution-amplification” with HBsAg coated in solid phase and help phage.The positive clone against HBsAg was characterized by ELISA assay and its P/N value was 2.415.The recombinant plasmid vectors from the positive clone was amplified by PCR with designed oligonucleotide primers of κ light chain.The κ chain PCR fragment was subcloned into plasmid pUC18 and sequenced.Results:The phage antibody against HBsAg was screened from a human immunoglouin phage displaying library.The pUC18-κ recombinant plasmid was constructed and sequenced.The gene size of κ chain is 643bp including variable region and constant region.Conclusion:According to the sequenceing and ELISA assay

we suggest that κ chain is a human anti-HBs-κ immunoglobuin.

关键词

人源性Ig轻链序列分析HBsAg

Keywords

Human Ig light chainSequencing analysisHBsAg

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