McLachlan A, Milich D R, Raney A K, Riggs M G, Hughes J L, Sorge J, Chisari F V
J Virol. 1987 Mar;61(3):683-92. doi: 10.1128/JVI.61.3.683-692.1987.
Amphotropic retroviral expression systems were used to synthesize hepatitis B virus surface antigen (HBsAg) and core antigen. The vectors permitted establishment of cell lines which expressed antigen from either the retroviral long terminal repeat or the mouse metallothionein-I promoter. HBsAgs were synthesized containing no pre-S sequences, pre-S(2) sequences alone, or pre-S(1) plus pre-S(2) sequences. Inclusion of pre-S(2) sequences did not affect the secretion or density of HBsAg particles but did reduce their mass by approximately 30%. Addition of pre-S(1) sequences almost completely abolished secretion of HBsAg and resulted in its localization in an aqueous-nonextractable pre- or early-Golgi cellular compartment. HBsAg was localized to the cytoplasm of the cell. This localization was unaffected by the presence of pre-S sequences in the antigen. Cell lines synthesizing hepatitis B antigens from core DNA fragments, containing or not containing precore sequences, secreted hepatitis B e antigen. However, the absence of precore DNA sequences resulted in additional synthesis of hepatitis core antigen, which was predominantly nuclear in localization.
双嗜性逆转录病毒表达系统被用于合成乙型肝炎病毒表面抗原(HBsAg)和核心抗原。这些载体能够建立细胞系,这些细胞系可从逆转录病毒长末端重复序列或小鼠金属硫蛋白-I启动子表达抗原。合成的HBsAg不含前S序列、仅含前S(2)序列或含前S(1)加前S(2)序列。包含前S(2)序列并不影响HBsAg颗粒的分泌或密度,但确实使其质量降低了约30%。添加前S(1)序列几乎完全消除了HBsAg的分泌,并导致其定位于水不可提取的前高尔基体或早期高尔基体细胞区室。HBsAg定位于细胞的细胞质中。这种定位不受抗原中前S序列存在与否的影响。从含有或不含有前核心序列的核心DNA片段合成乙型肝炎抗原的细胞系分泌乙型肝炎e抗原。然而,缺乏前核心DNA序列导致额外合成乙型肝炎核心抗原,其主要定位于细胞核。