Sako T, Tsuchida N
Nucleic Acids Res. 1983 Nov 25;11(22):7679-93. doi: 10.1093/nar/11.22.7679.
We have determined the entire nucleotide sequence of a 1,4-kilobase segment containing the staphylokinase gene, sak, molecularly cloned from the bacteriophage S phi-C genome of Staphylococcus aureus. The probable coding region is 489 base pairs long and these base pairs are translated into a polypeptide of 163 amino acid residues (Mr = 18,490) with a presumed signal sequence of 27 amino acid residues at the NH2-terminal end. In regions adjacent to the sak structural gene a possible promoter sequence and three possible terminator sequences for transcription were found about 100 base pairs upstream from the initiation codon and about 300, 400, and 500 base pairs downstream from the termination codon, respectively; they are active in an in vitro transcription system using Escherichia coli RNA polymerase. The immunoactive 18,500-dalton and 15,500-dalton proteins corresponding to a precursor form before secretion and a mature form after secretion of the sak gene products, respectively, were identified by the E. coli maxicell system.
我们已经确定了一个1.4千碱基片段的完整核苷酸序列,该片段包含从金黄色葡萄球菌噬菌体S phi-C基因组中分子克隆得到的葡萄球菌激酶基因sak。可能的编码区长度为489个碱基对,这些碱基对被翻译成一个由163个氨基酸残基组成的多肽(Mr = 18,490),在NH2末端有一个推测的由27个氨基酸残基组成的信号序列。在sak结构基因相邻区域,分别在起始密码子上游约100个碱基对处以及终止密码子下游约300、400和500个碱基对处发现了一个可能的启动子序列和三个可能的转录终止序列;它们在使用大肠杆菌RNA聚合酶的体外转录系统中具有活性。通过大肠杆菌大细胞系统鉴定出了与sak基因产物分泌前的前体形式和分泌后的成熟形式相对应的免疫活性18,500道尔顿和15,500道尔顿的蛋白质。