Qie Chun-hua, Qin Bo, Huang Ai-long, Liu Qi, Chen Guo-min
Viral Institute for Hepatitis, Key Laboratory of Molecular Biology on Infectious Diseases of Ministry of Education, Chongqing University of Medical Sciences, Chongqing 400010, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2006 Mar;22(2):141-3, 147.
To construct and express fusion gene encoding Hyper-IL-6 in mammalian cells.
Using geneSOEing method, human sIL-6R cDNA was fused with IL-6 cDNA by a linker rich in glycine through PCR technique and cloned into pIRES(2)-EGFP. The constructed plasmid was transfected into mammalian cells. The expression of fusion gene encoding Hyper-IL-6 was detected by GFP and RT-PCR and immunocytochemistry.
The recombinant plasmid pIRES-HIL-6 was successfully constructed and expressed in mammalian cells.
The success in construction and expression of human Hyper-IL-6 makes it possible to further study its biological functions.
在哺乳动物细胞中构建并表达编码Hyper-IL-6的融合基因。
采用基因拼接法，通过PCR技术将人sIL-6R cDNA与富含甘氨酸的接头连接到IL-6 cDNA上，并克隆到pIRES(2)-EGFP中。将构建好的质粒转染到哺乳动物细胞中。通过绿色荧光蛋白、逆转录聚合酶链反应和免疫细胞化学检测编码Hyper-IL-6的融合基因的表达。
成功构建了重组质粒pIRES-HIL-6，并在哺乳动物细胞中表达。
人Hyper-IL-6构建和表达的成功为进一步研究其生物学功能奠定了基础。
