摘要
将乙型肝炎病毒(HBV)ayw亚型Bam HI 1.5kb片段插入到载体pSV 2-dhfr Bgl Ⅱ位点,构建重组质粒。其正向插入的重组质粒pSV 2-DHBc-54转化L细胞之后,在细胞内测得乙型肝炎核心抗原(HBcAg)与e抗原(HBeAg),在含10％小牛血清的细胞培养液上清中仅测到HBeAg。此转化细胞用氨甲喋呤处理后,转化细胞表达HBcAg与HBeAg的量增加10倍以上。转化细胞系C-54-24传20代仍能稳定分泌HBeAg。转化细胞分泌的HBeAg,在含10％小牛血清的培养液中可在4℃稳定保存一年7个月以上。
In our preliminary report in l985,a, transformed mouse L cell line expressing HBcAg and HBeAg was established. This paper describes the detailed construction and expression data of this cell line. The BamH I 1.5 kb fragment of hepatitis B virus ayw subtype was inserted into the Bgl Ⅱ site of the, vector pSV2-dhfr and two recombinant plasmids of different orientations were constructed. HBcAg and HBeAg . were detected inside the transformed L cells transfected with the recombinant plasmid pSV2-DHBc-54 of correct orientation Only HBeAg was detected in the medium. The titers of HBcAg and HBeAg were amplified over 10-folds by methotrexate. HBeAg was stably secreted after 20 passages of the transformed L Cell line C-54-24. The HBeAg secreted by transformed L cell line stored at 4℃ was found to be stable for one year and seven months.
出处
《病毒学报》
CAS
CSCD
北大核心
1989年第3期201-210,共10页
Chinese Journal of Virology
关键词
乙肝病毒
核心抗原
E抗原
共转化
Hepatitis B core antigen Hepatitis, B e antigenCotrajjsformatiori Transformed mammalian cell line
