期刊论文

中文

解剖学杂志

1001-1633

2009

32

4

455-458

10.3969/j.issn.1001-1633.2009.04.008

国家自然科学基金（30270684） 湖南省教育厅一般项目

本校为第一机构

目的:构建真核表达重组体pEGFP-ZNF569,并鉴定其在哺乳动物COS-7细胞系能否正确表达.方法:把ZNF569基因ORF定向克隆入带有EGFP报告基因的真核表达载体,构建真核表达重组体pEGFP-ZNF569,脂质体介导将pEGFP-ZNF569转染入哺乳动物COS-7细胞,以一步法RT-PCR检测其在哺乳动物COS-7细胞中的表达情况.结果:双酶切及测序鉴定表明成功构建真核表达重组体pEGFP-ZNF569,并在COS-7细胞中成功地表达了融合荧光蛋白.RT-PCR检测显示RT-PCR扩增产物与ZNF569目的基因片段大小相符,确实源于重组质粒转录后的mRNA.结论:真核表达重组体pEGFP-ZNF569的成功构建及在体外真核细胞中有效表达,为进一步研究ZNF569在心发育...

Objective： To construct the recombinant plasmid of eukaryotic expression containing ZNF569 gene and transfect it into COS-7 cells to study the expression of ZNF569 and EGFP fusion proteins. Methods： ZNF569 gene was amplified from the plates by polymerase chain reaction （PCR）, and subcloned into an expression vector pEGFP-N1 containing the report gene of EGFP. After identification by sequencing and restrictive enzymes digestion, the recombinant plasmid pEGFP-ZNF569 was transfected into COS-7 cells using liposome. ZNF569 mRNA in COS-7 cells w...