中国寄生虫学与寄生虫病杂志 ›› 1992, Vol. 10 ›› Issue (2): 82-85.

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恶性疟杂合45肽抗原基因的化学合成及克隆

裘敏燕,闵永洁,王启松,钟雄林,陈仕荣,王昌才   

  1. 复旦大学遗传研究所; 复旦大学遗传研究所; 复旦大学遗传研究所; 第一军医大学分子生物学实验室; 第一军医大学分子生物学实验室; 第一军医大学分子生物学实验室
  • 收稿日期:2017-01-09 修回日期:2017-01-09 出版日期:1992-05-31 发布日期:2017-01-10

CHEMICAL SYNTHESIS AND CLONING OF PLASMODIUM FALCIPARUM 45 PEPTIDE ANTIGEN GENE

  • Received:2017-01-09 Revised:2017-01-09 Online:1992-05-31 Published:2017-01-10

摘要: 本文报道用固相亚磷酰胺法成功地合成了人恶性疟杂合抗原基因(Human Plasmodiumfalciparum antigen gene,HPFAG)。其全长为162bp,包含有三个裂殖子重复多肽(83kDa、55kDa、35kDa)和两个环子孢子表面重复多肽,以及两端的接头,共分成8个大小不同的片段合成。分离纯化后的片段连接成完整的双链基因,与P-Blue script重组,转化E.coli JM109,经原位杂交和酶切分析,筛选出重组体,又经DNA序列分析测定,证明与所设计基因完全相同。

关键词: 人恶性疟杂合肽抗原基因, DNA化学合成, 克隆

Abstract: We have synthesized a 162 bp gene of human Plasmodium falciparum hybrid peptide antigen by the solid-phase phosphoramidite method with ABI 381A DNA synthesizer.The gene encodes three fragments of the relative molecules 83kDa,55kDa and 35kDa merozoite-specific proteins and two CS repeats or four peptides.The gene with the designed two cohesive ends was divided into 8 fragments to be synthesized.All synthetic fragments were annealed and ligated with T4 DNA ligase to form double DNA chain.This synthetic gene was recombined with P-Blue script as vector and transformed into E.coli JM109.The positive recombinants were screened out by dot hybridization and enzyme analysis.The DNA sequence analysis showed that the synthesized human Plasmodium falciparum hybrid peptide antigen gene was identical with the designed one.(Figs.1-4)