sgp130 cDNA在痘苗病毒载体系统中的表达

EXPRESSION OF SOLUBLE gp130 GENE IN VACCINIA VIRUS SYSTEM

ｇｐ１３０是一种分子量为１３０ｋＤ的细胞膜糖蛋白，是ＩＬ－６等多种细胞因子共用的信号传递分子。本研究将含ｇｐ１３０胞外区全长的可溶性片段的基因（ｓｇｐ１３０ｃＤＮＡ）构建到痘苗病毒载体中，并与野生型痘苗病毒在ＴＫ－１４３细胞中发生同源重组，用ＢｕｄＲ和Ｘ－ｇａｌ双重选择，得到带有人ｓｇｐ１３０的重组痘苗病毒（Ｖｓｇｐ１３０）。采用ＩＤＡ和ＷｅｓｔｅｒｎＢｌｏｔｉｎｇ法证实：感染Ｖｓｇｐ１３０的ＴＫ－１４３细胞上清中有较强的ｓｇｐ１３０表达，表达产物分子量为１００ｋＤ左右。

gp130 is a cell membrane glycoprotein of 130 kDa.Some cytokines such as IL 6,ciliary neurotro phic factor(CNTF),oncostatin M(OM)and leukemia inhibitory factor(LIF) share the gp130 as a common signal transducer.We have constructed soluble gp130(sgp130) recombinant expression vector by inserting the sgp130 cDNA into the vaccinia virus expression plasmid pJSA1175.This recombinant plasmid was then used for homologous recombination with the wild type vaccinia virus genome in TK -143 cells.sgp130 recombinant vaccinia virus(Vsgp130) was then obtained by the BudR and X gal double sellections.Immunodot assay and Western blotting analysis showed that the recombinant vaccinia virus could express sgp130 into the supernatant of TK -143 cells and the molecular weight of expression product was about 100 kDa.