摘要
人白细胞介素 1 2 (hIL 1 2 )是细胞介导免疫发生的关键调节因子 ,也是目前发现的唯一的异源二聚体细胞因子 ,由P35和P40两个亚基经二硫键连接而成。利用DNA重组技术 ,分别构建了含hIL 1 2 p35基因和 p40基因的重组转移载体质粒pAcAB3 p35和pAcAB3 p40。将两个重组转移载体分别与致死缺陷型线性化苜蓿丫纹液蛾核型多角体病毒 (AcNPVBaculoGoldLinearizedBaculovirus)基因DNA共转染昆虫细胞 ,构建出遗传稳定的重组病毒AcNPV OCC hIL 1 2 (p35)与AcNPV OCC- hIL 1 2 (p40 )。将两种病毒分别感染Sf9细胞 ,取细胞培养物上清和细胞裂解物上清进行SDS PAGE和Westernblot检测 ,结果显示hIL 1 2p35和 p40两基因均在昆虫细胞中获得表达 ,且能分泌至胞外。表达产物具有免疫原性。
Human Interleukin\|12 (hIL\|12) is a key immunoregulatory cytokine and the only heterodimeric cytokine found by far. It is a disulfide\|linked heterodimer composed of a 35kD light chain (P35) and a 40kD heavy chain (P40). Two recombinant transfer vector plasmids pAcAB3\| p 35 (including hIL\|12 p 35 gene) and pAcAB3\| p 40 (including hIL\|12 p 40 gene) were constucted and co\|transfected the insect cells Sf9 with AcNPV BaculoGold Linearized Baculovirus DNA. The obtained recombinant viruses, AcNPV\|OCC\+-\|hiL\|12 (\%p\%35) and AcNPV\|OCC\+-\|hIL\|12 ( p40 ), were used to infect the Sf9 cells separately, and the supernatants of culture and cell lysate were examined by using SDS\|PAGE and Western blot. Results showed that both hIL\|12 p35 and p40 gene were successfully expressed in insect cells and secreted. The expression productions had immunogenicity.
出处
《微生物学报》
CAS
CSCD
北大核心
2001年第1期35-42,共8页
Acta Microbiologica Sinica
基金
广东省自然科学基金资助项目!( 960 0 59)&&
关键词
人白细胞介素
杆状病毒表达系统
表达
Human interleukin\|12 (hIL\|12), Baculovirus expression system, Expressio(
