胞红蛋白在CHO细胞中的表达及其抗氧化活性研究  

Expression and Antioxidant Activity of Cytoglobin in CHO Cell

作　　者：李世飞[1] 贾晓健[1] 吴静[1] 钱杰[1] 陶伟娟[1] 吴梧桐[1] 张玉彬[1] 

机构地区：[1]中国药科大学生物化学教研室,江苏南京210009

出　　处：《药物生物技术》2011年第4期296-299,共4页Pharmaceutical Biotechnology

基　　金：南京市优秀留学回国人员资助项目(S102003)

摘　　要：构建胞红蛋白(Cytoglobin,CYGB)真核表达载体,并在CHO细胞中进行表达。通过PCR技术扩增了CYGB基因序列,并将CYGB基因序列定向插入pcDNA3.1(+)表达载体中,构建了pcDNA3.1-CYGB真核表达载体。将重组表达载体pcDNA3.1-CYGB转染CHO细胞,经G418筛选出阳性细胞株。培养、收集细胞,提取细胞总蛋白,经Western Bloting鉴定证明CYGB表达。通过H2O2(800μmol/L)诱导的氧化应激损伤实验,检验CYGB的抗氧化活性,与对照组相比,CYGB表达细胞株提高了细胞内超氧化物酶(SOD)的活力,显著的提高了细胞在H2O2诱导的氧化应激下的存活率。研究结果表明,真核表达载体pcDNA3.1-CYGB构建成功,CYGB在真核细胞CHO中成功表达。The eukaryotic expression vector of Cytoglobin （pcDNA3. 1 - CYGB） was constructed to express Cytoglobin in mammalian CHO cell. CYGB gene was amplified by using PCR and inserted into pcDNA3. 1 （＋） plasmid. The recombinant plasmid was transfected into CHO cell and then the transfected cells were screened with G418. The total protein was extracted from positive clones and Western Bloting was used to identify the expression of CYGB. Antioxidant activity of CYGB was analyzed by cell survival assay of H2 02 induced oxidative stress damage. The data showed that over - expression of CYGB enhanced SOD （superoxide dismutase） activity in CHO cell and increased cell survival. Therefore, the recombinant vector pcDNA3.1 - CYGB was correctly constructed and CYGB was expressed successfully in CHO cell. CYGB overexpression can enhance antioxidant activity and protect cell from death induced by H2O2.

关 键 词：胞红蛋白 CHO细胞 表达 抗氧化活性 

分 类 号：Q51[生物学—生物化学] Q786