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Granulysin真核表达载体的构建与鉴定

Construction of Eukaryotic Expression Vector of Granulysin and its DNA Sequencing
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摘要 目的:构建颗粒溶解素(granulysin)cDNA真核表达载体。方法:采用逆转录聚合酶链反应(RT-PCR)技术从健康志愿者外周血单个核细胞(PBMC)中扩增出颗粒溶解素蛋白cDNA,经限制性核酸内切酶消化后,插入真核表达载体pcDNA3.1(-)的相应酶切位点,将此重组质粒进行序列测定,并将重组质粒转染COS7细胞,进行基因表达鉴定。结果:重组质粒插入基因序列测定证实为颗粒溶解素cDNA,并能在COS7细胞稳定表达。结论:成功构建颗粒溶解素真核表达载体,为进一步研究其抗结核作用奠定了基础。 Objective: To construct eukaryotic expression vector of Granulysin. Methods: Granulysin cDNA was amplified from PBMC of healthy volunteer by RT-PCR, and correctly inserted into corresponding sites of eukaryotic expression vector pcDNA3. 1 (-) after restriction endonuclease digestion, and the recombinant plasmid was transfected into COS7 cells, gene expression was determined by RT-PCR. Results: The gene fragment inserted into the vector pcDNA3.1(-) was confirmed by nucleotide sequencing, and the mRNA of the recombinant plasmid was successfully expressed in mammalian cells. Conclusion: The successful construction of recombinant eukaryotic expression vector based on the cDNA of granulysin, provides the experiment basis for further researching.
出处 《武汉大学学报(医学版)》 CAS 2005年第6期804-807,共4页 Medical Journal of Wuhan University
基金 湖北省科技厅重大课题资助项目(301121028)
关键词 颗粒溶解素 CDNA 真核表达载体 Granulysin cDNA Eukaryotic Expression Vector
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