Kotani H, Ishizaki Y, Hiraoka N, Obayashi A
Nucleic Acids Res. 1987 Mar 25;15(6):2653-64. doi: 10.1093/nar/15.6.2653.
The coding region of the gene for bacteriophage SP6 RNA polymerase was cloned into pBR322, and its entire nucleotide sequence was deduced. The predicted amino acid sequence for the polymerase consists of 874 amino acid residues with a total molecular weight of 98,561 daltons. Comparison of the amino acid sequence with that of T7 RNA polymerase reveals that regions with partial homology are present along the sequence. The coding region of SP6 RNA polymerase was inserted into an E. coli expression vector. The polymerase gene was efficiently expressed in E. coli cells, and the enzymatic properties of the expressed polymerase were very similar to those of the enzyme synthesized in SP6 phage-infected Salmonella typhimurium cells.
将噬菌体SP6 RNA聚合酶基因的编码区克隆到pBR322中，并推导其完整的核苷酸序列。该聚合酶预测的氨基酸序列由874个氨基酸残基组成，总分子量为98,561道尔顿。将该氨基酸序列与T7 RNA聚合酶的序列进行比较，发现沿序列存在部分同源区域。将SP6 RNA聚合酶的编码区插入大肠杆菌表达载体中。该聚合酶基因在大肠杆菌细胞中高效表达，表达的聚合酶的酶学性质与在SP6噬菌体感染的鼠伤寒沙门氏菌细胞中合成的酶非常相似。
