人granzyme B基因在HeLa细胞中的表达
基因表达,,granzyme,B;基因表达;HeLa细胞,人granzymeB基因在HeLa细胞中的表达,0引言,1材料和方法,2结果,3讨论,参考文献:
关键词: granzyme B;基因表达;HeLa细胞摘 要:目的 构建携带人granzyme B基因的真核表达载体,并将其在HeLa细胞中表达. 方法 用反转录PCR获取人granzyme B全长cDNA序列,将其活性型序列克隆进pcD-NA3重组表达质粒.脂质体法转染HeLa细胞,间接免疫荧光检测目的蛋白表达,HE染色观察其对转染HeLa细胞形态的影响. 结果 成功获得了野生型granzyme B cDNA,构建了编码活性型granzyme B的真核表达载体pcDNA3-G.转染HeLa细胞后观察到目的蛋白表达,转染细胞形态发生变化,出现多核大细胞及固缩小细胞. 结论 活性型granzyme B哺乳动物表达系统的建立,为进一步研究granzyme B的生物学效应奠定了基础.
Expression of human granzyme B gene in HeLa cells
ZHAO Jing XU Yan-Ming JIA Lin-Tao YU Cui-Juan CHEN Lei HUANG Wei-Quan WANG Cheng-Ji
YANG An-Gang
1 Department of Biochemistry and Molecular Biolo-gy,
2 Department of Histology and Embryology,Faculty of Preclinical Medicine,Fourth Military Medical University,Xi'an710033,China
Keywords:granzyme B;gene expression;HeLa cells
Abstract:AIM To construct eukaryotic expression vector carrying human granzyme B gene and express it in HeLa cells.METHODS RT-PCR was used to obtain human granzyme B cDNA.Then the recombinant expression vector pcDNA3-G was constructed by cloning active ganzyme B into eukaryotic expression vector pcDNA3.After transfection into HeLa cells through lipofectamine2000 ......
您现在查看是摘要页,全文长 9551 字符。