摘要
根据细胞因子协同作用的特点,采用重组DNA技术构建了人干扰素(IFN)α2b 胸腺肽(THY)α1融合基因,克隆到pBacPAK8上,获得重组转移载体pBacPAK IFN THY.与线形化Bm BacPAK6病毒基因组DNA共转染家蚕细胞,经过体内重组,筛选到重组病毒Bm BacPAK IFN THY.将Bm BacPAK IFN THY感染家蚕细胞进行表达.DNA印迹证明IFN THY已插入Bm BacPAK6中(4kb左右的杂交带);SDS 聚丙烯酰胺凝胶电泳、蛋白质印迹证明IFN THY在家蚕细胞中得到了表达(分子质量为 2 3ku左右),且具有IFN蛋白的免疫原性;微量细胞病变抑制法和玫瑰花结法显示 96h的表达产物IFN活性为 3 72× 10 4U/ml,12 0h表达产物IFN活性为 3 10× 10 5U/ml,4 8～ 72h表达产物IFN活性较低;4 8～ 12 0h表达产物的玫瑰花结形成率均在 10 %以上.结果表明融合基因在家蚕细胞中得到了高效表达,表达的融合蛋白具有IFN α2b和THY
The baculovirus shuttle vector , pBacPAK IFN THY was constructed which contains the genes of IFN α2b and THY α1. Constructed vector was coinfected with linear Bm BacPAK6 DNA into BmN cells. The recombinant virus was screened and plaque purified. The BmN cells were infected with the recombinant virus. The results showed that the protein was successfully prepared and its bioactivities were testified by WISH VSV system and E RFC. Results indicated that the expressed fusion protein have bioactivities of both IFN α2b and THY α1.
出处
《生物化学与生物物理进展》
SCIE
CAS
CSCD
北大核心
2003年第5期803-807,共5页
Progress In Biochemistry and Biophysics
