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郑长青
白细胞介素-2,DNa,重组,诱变
【摘要】 目的 构建基因变构IL-2( 88 N→R)的重组克隆。方法 将正常人的扁桃体细胞经PHA刺激后，获得cDNA文库，并以此为模板，经套式PCR扩增得IL-2的基因编码序列。测序确认正确后，设计含有突变位点的引物，经PCR定点诱变技术获得变构IL-2克隆并进行序列测定。结果 IL-2基因定点诱变成功，并获得了基因变构IL-2( 88 N→R)重组克隆。结论 IL-2重组基因变构克隆的构建为进一步在原核和真核细胞中表达以及研究制备低毒、高效的新型基因变构IL-2奠定了基础。
关键词 白细胞介素-2 DNA 重组 诱变
【文献标识码】 A 【文章编号】 1606-8106(2003)09-1286-03
Clone construction of a site-specific gene
mutant of the human interleukin-2
Zheng Changqing，Feng Guoji，Qian Dongmeng，et al.
Naval No.401Hospital of PLA，Qingdao266021.
【Abstract】 Objective To construct clones of a site-specific mutagenesis( 88 N→R)of human interleukin-2(IL-2)gene.Methods Nested-PCR was employed to amplify gene coding for IL-2from the PHA-stimulated human tonsil cell cDNA.Then the specific primers containing the mutated base were synthesized to obtain the target fragments by the means of recombinant PCR.The amplified fragments were cloned to pGEM-T Easy vector and transˉformed into E.coli.JM109.We got the positive clone by blue/white screening.Results The mutated IL-2gene clone was obtained s ......
关键词 白细胞介素-2 DNA 重组 诱变
【文献标识码】 A 【文章编号】 1606-8106(2003)09-1286-03
Clone construction of a site-specific gene
mutant of the human interleukin-2
Zheng Changqing，Feng Guoji，Qian Dongmeng，et al.
Naval No.401Hospital of PLA，Qingdao266021.
【Abstract】 Objective To construct clones of a site-specific mutagenesis( 88 N→R)of human interleukin-2(IL-2)gene.Methods Nested-PCR was employed to amplify gene coding for IL-2from the PHA-stimulated human tonsil cell cDNA.Then the specific primers containing the mutated base were synthesized to obtain the target fragments by the means of recombinant PCR.The amplified fragments were cloned to pGEM-T Easy vector and transˉformed into E.coli.JM109.We got the positive clone by blue/white screening.Results The mutated IL-2gene clone was obtained s ......