摘要
将乙肝病毒表面抗原（ＨＢｓＡｇ） 主蛋白的编码基因Ｓ与人ＧＭＣＳＦ 基因融合后， 插入真核表达质粒ｐｃＤＮＡ１ 中， 经质粒酶切鉴定及ＤＮＡ 序列分析证明， 目的基因（１． １ｋｂ） 插入ｐｃＤＮＡ１ 的ＣＭＶ 启动子下游。以获得的重组质粒ｐＳＧＭ 转染Ｌ９２９ 细胞， 经ＲＴＰＣＲ 及细胞原位杂交证实了目的基因的转录。通过免疫组化技术， 利用抗ＨＢｓ 和ＧＭＣＳＦ抗体， 检出目的基因在Ｌ９２９ 细胞中的表达。该结果为用乙型肝炎病毒的基因免疫打下了基础。
HBV S gene encoding major envelope protein was fused with human GM CSF gene and inserted into eukaryotic expression plasmid pcDNA1. Enzyme digestion and DNA sequencing analysis showed that the 1.1 Kb target gene was inserted into downstream of the CMV promoter. Then the recombinant plasmid pSGM was transfected into L929 cells. The gene transcription was confirmed by RT PCR and cell in situ hybridization and the gene products was demonstrated by immunohistochemical staining with anti HBs and anti GM CSF antibody. Above results lay the foundation for the gene immunization of HBV.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
1999年第4期259-262,共4页
Chinese Journal of Cellular and Molecular Immunology
