Thummel C S, Burgess T L, Tjian R
J Virol. 1981 Feb;37(2):683-97. doi: 10.1128/JVI.37.2.683-697.1981.
We constructed a series of bacterial plasmids which contained the Escherichia coli lac promoter fused to a simian virus 40 restriction fragment coding for small t antigen. These plasmids expressed different levels of intact viral protein depending on the length of the constructed ribosome binding site. Small t antigen synthesized by the most efficient producer, HP1, constituted 0.5 to 1% of the total cellular protein. On the basis of extensive characterization by immunoprecipitation, gel electrophoresis, isoelectric focusing, tryptic fingerprint analysis, and chromatographic properties, this plasmid-encoded protein was virtually identical to authentic simian virus 40 small t antigen. Partial purification of the HP1-encoded and authentic small t antigens revealed the presence of both monomeric and multimeric forms.
我们构建了一系列细菌质粒，这些质粒包含与编码小t抗原的猿猴病毒40限制片段融合的大肠杆菌乳糖启动子。根据构建的核糖体结合位点的长度，这些质粒表达不同水平的完整病毒蛋白。由最高效的生产者HP1合成的小t抗原占细胞总蛋白的0.5%至1%。通过免疫沉淀、凝胶电泳、等电聚焦、胰蛋白酶指纹分析和色谱特性进行广泛表征，这种质粒编码的蛋白与 authentic 猿猴病毒40小t抗原几乎相同。对HP1编码的和 authentic 小t抗原进行部分纯化，发现同时存在单体和多聚体形式。
