摘要
研究了由含乙型肝炎病毒C基因与前-C基因的重组质粒,转化小鼠L细胞所分泌的乙型肝炎病毒e抗原(HBeAg)的性质。此种HBeAg有与牛血清蛋白非特异性结合的性质,在牛血清培养基中可能是与α和β球蛋白结合。结合的HBeAg,分子量约60～80KD,在50硫酸铵中可完全析出.经SDS-PAGE解离后,Western blot证明,牛血清培养基中的HBeAg的分子量为28KD、24KD、22KD和19KD4种。无血清培养基中的HBeAg以聚合体形式存在,在66.6％硫酸铵中方可完全析出。经SDS-PAGE解离后,Western blot证明,HBeAg的分子量为56KD和28KD。28KD分子量与由DNA序列推算的前-C和C基因编码的理论值一致。单克隆抗体检测证明,转化细胞分泌的HBeAg具有HBeAg/a和HBeAg/b抗原决定簇。在免疫扩散试验中,转化细胞分泌的HBeAg与人血清抗-HBe形成一条沉淀线,与人血清HBeAg近抗体侧的沉淀线端端相连,但与标准血清EK-3(抗-HBel和抗-HBe2)和EK-1(抗-HBe2和R-HBe3)在EIA试验中均呈阳性反应。
We have studied the properties of HBeAg secreted by L cells transformed with recombinant plasmid containing prscore and core genes of the hepatitis B virus. The HBeAg secreted by transformed mammalian cells in 10% calf serum medium was found to be non-specifically associated with calf serum proteins, which may be the α1 and (3 globulins. The molecular weight of HBeAg associated with serum proteins was about 60-80KD. It was completely precipitated by 50% ammonium sulfate. After disassociation by SDS-PAGE, the molecular weights of HBeAg in 10% calf serum medium as identified by Western blot were found to be 28KD,24KD, 22KD and 19KD. On the contrary,HBeAg in serum-free medium was completely precipitated only by 66.6% ammonium sulfate. Its molecular weights as identified by Western blot were found to be 56KD and 28KD. 56KD HBeAg may be represent the dimer. The disassociated 28KD HBeAg is equivalent in molecular weight to the theoretical value calculated from the sequence of precore and core gene, while the 24KD, 22KD and 19KD HBeAg may represent different break-down products. HBeAg secreted by transformed L cells possessed both HBeAg/a and HBeAg/b determinants as identified by monoclonal antibodies.
出处
《病毒学报》
CAS
CSCD
北大核心
1990年第1期1-11,共11页
Chinese Journal of Virology
