中华皮肤科杂志 ›› 1996, Vol. 29 ›› Issue (1): 31-34.

• 论著 • 上一篇    下一篇

160000类天疱疮抗原片段cDNA克隆的建立及测序

李冠群, 朱学骏   

  1. 北京医科大学第一医院皮肤科, 100034
  • 收稿日期:1995-04-25 修回日期:1995-08-02 出版日期:1996-02-15 发布日期:1996-02-15
  • 基金资助:
    卫生部基金资助课题

Construction and Sequencing of cDNA Clone of Bullous Pemphigoid 160000 Antigen Fragment

Li Guanqun, Zhu Xuejun   

  1. Department of Dermatology, First Teaching Hospital, Beijing Medical University, Beijing 100034
  • Received:1995-04-25 Revised:1995-08-02 Online:1996-02-15 Published:1996-02-15

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摘要： 大疱性类天疱疮(BP)存在两个自身抗原:BPAGl(220000～240000)和BPAG2(160000～180000),二者为不同的基因产物。为了研究BP的分子生物学特性,我们通过PCR技术建立了BPAG2片段的cDNA克隆。首先根据已知序列合成了特异引物,下游引物导入Hind Ⅲ酶切位点。然后从人角朊细胞cDNA文库中扩增得到BPAG2氨基端588bp的cDNA片段,经NcoⅠ和HindⅢ酶切位点连入质粒载体pKPL-3a,最后转化受体菌pop2136。我们筛选得到了一个重组阳性克隆,经酶切和测序分析证实重组克隆的插入片段为BPAG2编码细胞内氨基端非胶原区的片段。为进一步研究其体外表达和抗原特性奠定了基础。

关键词: 类天疱疮,大疱性, 抗原, cDNA

Abstract: Bullous pemphigoid has two autoantigens:BPAG1 (220 000～240 000) and BPAG2 (160 000～180 000),which are distinct gene products.In this paper we constructed a cDNA clone encoding BPAG2 fragment by PCR to study the molecular characters of BPAG2.First we synthesized specific primers,and a Hind Ⅲ restriction enzyme site was added to the downstream primer.Then a 588bp at BPAG2 amino terminal was amplified from human keratinoeyte cDNA library.This PCR products were cloned into a plasmid pKPL-3a digested with Nco Ⅰ and Hind Ⅲ and then transformed E.coli. pop2136.We screened out a positive recombinant clone,and confirmed that the insert in this recombi-nant encoded the amino-terminal non-collagenous domain of BPAG2 by restriction enzymes digestion and DNA sequencing.

Key words: Pemphigoid,bullous, Antigens, cDNA