摘要
将人肿瘤坏死因子α（Ｔｕｍｏｒ Ｎｅｃｒｏｓｉｓ ＦａｃｔｏｒαＴＮＦ－α）ｃＤＮＡ片段克隆到转移载体ｐＡｃ６１０上，然后与苜蓿尺蠖核型多角体病毒（Ａｕｔｏｇｒｏｐｈａ ｃａｌｉｆｏｒｎｉｃａ Ｎｕｃｌｅａｒ Ｐｏｌｙｈｅｄｒｉｎ Ｖｉｒｕｓ，ＡｃＮＰＶ）ＣＮＡ共转染草地贪夜蛾细胞（Ｓｐｏｄｏｐｔｅｒｓ ｆｒｕｇｉｐｅｒｄａ）Ｓｆ２１，获得含ｈＴＮＦ－α为１．０×１．＾４ｕ／１０＾６细胞：同时。
A cDNA fragment coding human tumor necrosis factor a (hTNF-α) was inserted into the vector pAC610. The Sf21 cells (Spodopters frugiperda 21) were co-trans-fected with pAC610-hTNF-a and AcNPV DNA (Autographa californica Nuclear Poly-hedrin Virus DNA). The resultant recombinant virus was screened by dot hybridization and purified by plaque formation technique. TNF-α activity was measured by L929 cyto-toxic assay. The highest expression level, 5.2 × 104u/106 cells, was given at 72 hours after infection and with 6% FBS in medium. ELISA tests showed that the product expressed in hTNF-α.
出处
《生物工程学报》
CAS
CSCD
北大核心
1995年第1期73-76,共4页
Chinese Journal of Biotechnology
关键词
肿瘤坏死因子
Α基因
基因表达
昆虫细胞
Tumor necrosis factor a. baculovirus. Spodoptera frugiperda 21 cells, gene expression
