可溶性CD14基因在COS-7细胞的表达  

Transient expression of soluble CD14 gene in COS-7 cells

作　　者：王胜朝[1] 荫俊[2] 史俊南[1] 白洁[3] 王威[2] 宋伟[2] 

机构地区：[1]第四军医大学口腔医学院,陕西西安710032 [2]军事医学科学院微生物流行病研究所,北京100071 [3]白求恩医科大学第二临床医学院,吉林长春130021

出　　处：《牙体牙髓牙周病学杂志》2002年第9期463-465,共3页Chinese Journal of Conservative Dentistry

基　　金：国家自然科学基金项目 (3 9870 3 16)

摘　　要：目的 :构建重组可溶性CD14真核表达载体 ,用瞬时系统表达目的基因。方法 :将可溶性CD14基因定向克隆入真核表达质粒载体 pcDNA3.1+;DEAE -葡聚糖法转染COS - 7细胞 ,72h收集培养上清 ,Westernblot鉴定表达产物 ,ELISA检测表达水平。结果 :酶切鉴定表明重组表达载体构建成功 ;SDS -PAGE显示转染细胞可表达一分子量 5 2 .6× 10 3 u (5 3kDa)的蛋白 ;Westernblot显示 5 2 .6× 10 3 u (5 3kDa)处有免疫阳性蛋白带 ;ELISA检测表明 :转染 72h目的蛋白可达 2 .1μg/mL。 结论 :重组可溶性CD14基因可以在COS -AIM:To express the soluble CD14 gene in COS-7 cells. METHODS: Soluble CD14 gene was subcloned into mammalian expression vector pcDNA3.1+, the recombined plasmids were transfected into COS-7 cells using DEAE-dextran kit for transient expression. Western-blot analysis and ELISA assay were used to examine the supernatant. RESULTS: The constructed vectors were confirmed by digestion with restriction enzyme. Western-blot analysis of culture supernatant found a immuno-reaction positive band with molecular weight of 53kDa. ELISA assay revealed that the amount of rsCD14 in the serum-free medium was about 1.5 μg/mL.CONCLUSION: Recombined soluble CD14 gene can be expressed effectively in COS-7 cells.

关 键 词：CD14 C0S-7 真核表达 Gˉ菌感染 Gˉ菌内毒素脂多糖 血清 髓细胞 

分 类 号：R780.2[医药卫生—口腔医学]