摘要
含信号肽序列的hG-CSFcDNA基因克隆于M13mp18中,测序表明其基因序列与高活性hG-CSFcDNA一致。将hG-CSF基因插入家蚕核型多角体病毒(BmNPV)转移载体pBE284,经与野生型病毒DNA共转染家蚕细胞后,通过体内同源重组的方式构建了重组病毒BmNPV-G-CSF,Southern印迹杂交表明重组病毒DNA中含G-CSF基因片段。重组病毒感染单层BmN细胞后第四天表达量达1. 2×106CFU/ml培养液,Western-blot分析可见分子量为19×103左右一条带。家蚕幼虫感染重组病毒后第四天表达量达1.4×107CFU/ml血淋巴(hemolymph)。
The cDNA of hG-CSF containing signal sequence was cloned into M13mp1 8 vector , and the sequence analysis showed that the cloned fragment was the high active form of hG-CSF. Insertion of the hG-CSF. cDNA into BmNPV transfer vector pBE284 , formed a recombinant transfer vector pBE284-G. BmN cell was cotransfected with wild type viral DNA and pBE284-G. The recombinant virus BmG-CSF was obtained by homologous recombination in vivo Southern-hybridization analysis suggests that the recombinant viral DNA contains hG-CSF cDNA fragrnent. The biological activity of G-GSF was 1 . 2 × 1 06CFU /ml at day 4 in the supernatant of BmN cell culture infected with BmG-CSF and was 1 . 4× 107CFU /ml at day 4 in the hemolymph of silkworm larvae infected with BmG-CSF.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
1996年第1期29-32,共4页
Chinese Journal of Microbiology and Immunology
关键词
G-CSF
序列分析
家蚕
基因表达
生物活性
Human granulocyte colony-stimulating factor
Sequence analysis
Silkworm Gene expression
Biological activity