摘要
利用放射免疫分析（ＲＩＡ）证实了含金鱼生长激素ⅠｃＤＮＡ的重组病毒在感染细胞９６ｈ后所表达的生长激素达到最高水平，平均每１０５个细胞可分泌金鱼生长激素Ⅰ最高达２８８０７ｎｇ；平均每克干虫可产生金鱼生长激素Ⅰ９２５３μｇ。从感染重组病毒的无血清细胞培养基中纯化生长激素Ⅰ，平均每毫升培养基可纯化具免疫活性纯度９５％以上的金鱼生长激素Ⅰ达６６４２２４ｎｇ。纯化后蛋白的Ｎ末端首２０个氨基酸序列测定的结果表明其信号肽得到了准确的切割。从而保障了表达蛋白的生物活性。
The expression level in vivo and in vitro were quantified using RIA.Average of 10 5 Sf9 cells may secret gfGHI into medium reaching level of 288 07ng in 96 h pi(post infection);The expression level of gfGHI in larvae may reach to 925 3mg per gram of dry larvae.The gfGHI was purified from the serum free medium(SFM) of cultured Sf 9 cells which infected with recombinant viruses containing gfGHI gene.666.224ng of gfGHI whose purity higher than 95% was purified from per minilitre SFM medium.The results of protein sequencing of N terminal of purified gfGHI proved that the signal peptide were cleaved accurately.These results,taken together,suggest that gfGHI were highly expressed successfully using baculovirus expression system and the expressed gfGHI was modified by this system.
出处
《生物工程学报》
CAS
CSCD
北大核心
1998年第4期419-423,共5页
Chinese Journal of Biotechnology
基金
香港RGC资助
关键词
杆状病毒
表达载体
金鱼生长激素
基因
表达
Baculovirus expression vector system,goldfish growth hormone gene,RIA,purification
