Baek M C, Kim S K, Kim D H, Kim B K, Choi E C
College of Pharmacy, Seoul National University, Korea.
Microbiol Immunol. 1996;40(8):531-7. doi: 10.1111/j.1348-0421.1996.tb01105.x.
A gene-encoding arylsulfate sulfotransferase (ASST) was cloned from a Klebsiella K-36 genomic library. ASST transfers a sulfate group from phenolic sulfate esters to a phenolic acceptor substrate. The gene, designated astA, was subcloned into vector pGEM3Zf(-) and sequenced. Recombinant clone-harbouring astA was directly identified using a fluorescent product. The nucleotide sequencing revealed an open reading frame (ORF) of 2,082 bp encoding a protein of 694 amino acids with a secretory signal sequence. A protein of similar size was visualized after in vitro transcription and translation using a plasmid carrying the cloned 3.1-kb fragment as a template. The N-terminal amino acid sequence of the purified processed protein was found to be identical to that predicted from the gene sequence. When searching the database for astA nucleotide or its deduced amino acid sequence, no significant homology to any sequence was found.
从肺炎克雷伯菌K-36基因组文库中克隆出一个编码芳基硫酸酯硫酸转移酶（ASST）的基因。ASST将硫酸基团从酚硫酸酯转移至酚类受体底物。该基因命名为astA，被亚克隆至载体pGEM3Zf(-)并进行测序。携带astA的重组克隆通过荧光产物直接鉴定。核苷酸测序揭示了一个2082 bp的开放阅读框（ORF），编码一个含有分泌信号序列的694个氨基酸的蛋白质。使用携带克隆的3.1 kb片段的质粒作为模板进行体外转录和翻译后，可见到一个大小相似的蛋白质。纯化的加工后蛋白质的N端氨基酸序列与基因序列预测的序列相同。在数据库中搜索astA核苷酸或其推导的氨基酸序列时，未发现与任何序列有显著同源性。
