【摘要】 将hIGF-I基因克隆进原核表达载体pET-28a(+),在E.coli中进行了融合表达,West-ern blotting显示在26 kD附近有一条特异条带。将hIGF-I基因克隆进pBacPAK-8,获得了杆状病毒转移载体pBacPAK-8-IGF-I,在脂质体的介导下,与线性化的家蚕杆状病毒共转染家蚕培养细胞Bm-N,经空斑筛选,PCR检测,获得了重组病毒Bm-Bac-hIGF-Ⅰ。SDS-PAGE检测表明,在感染重组病毒后,家蚕幼虫血淋巴中可以检测到一条分子量约为7.5 kD的特异性条带,ELISA检测表达量达4.51μg/mL蚕血淋巴。更多还原

【Abstract】 A 588bp cDNA gene sequence(hIGF-I) of recombinant human insulin-like growth factor(hIGF-I) was cloned into pET-28a(+).The hIGF[WTBZ]-I expression was performed in E.coli.SDS-PAGE and Western blotting suggests that the hIGF-I has the molecular weight of about 26 kD.The hIGF-I was cloned into pBacPAK-His8 to form pBacPAK-His-hIGF-I plasmid.Recombinant virus Bm-Bac-hIGF-I was generated by co-infection into BmN cells with the transfer plasmid pBacPAK-His-hIGF-I and the modified Bombyx mori nuclear polyhedrosis virus Bm-BacPAK8 genomic DNA.SDS-PAGE shows that recombinant protein expressed in larvae of silkworm is about 7.5 kD in molecular weight,and expression level of 4.51μg per milliliter hemolymph was detected by ELISA.更多还原