期刊论文

中文

实用预防医学

1006-3110

2005

12

6

1257-1260

10.3969/j.issn.1006-3110.2005.06.001

30270684:中国科学院项目（非规范项目） 02JJYB004:湖南省***科学基金

本校为第一机构

目的构建真核表达重组体pEGFP-M-CSF,并鉴定其在小鼠成纤维细胞NIH3T3细胞能否正确表达.方法PCR扩增M-CSF基因,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-M-CSF,脂质体介导将pEGFP-M-CSF转染入NIH3T3细胞,以一步法RT-PCR检测其在NIH3T3细胞中的表达情况.结果双酶切及测序鉴定证明成功构建真核表达重组体pEGFP-M-CSF,并在NIH3T3细胞中成功地表达了融合荧光蛋白,RT-PCR检测结果显示RT-PCR扩增产物与M-CSF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-M-CSF的成功构建及在体外真核细胞中有效表达,为进一步研究M-CSF在真核细胞中的信号调控...

Objective To construct the recombinant plasmid of eukaryotic expression vector containing M-CSF gene and transfect it into the NIH3T3 cells to study whether resulting in the expression of M- CSF and EGFP fusion proteins. Methods M-CSF gene was amplified by polymerase chain reaction （PCR）, and subcloned into an expression vector pEGFP-C1 containing the report gene of EGFP. After identification by sequencing and restrictive enzymes digestion, the recombinant plasmid pEGFP- M- CSF was transfected into NIH3T3 cells by using the liposome technique...