偶氮染料脱色菌株AZR偶氮还原酶基因的克隆及其序列分析

摘要

利用16S rRNA鉴定偶氮染料脱色菌株AZR属于葡萄球菌属(Staphylococcus)的科氏葡萄球菌(Staphylococcus cohnii),根据GenBank上登录的葡萄球菌属的3个偶氮还原酶基因序列设计扩增引物,从菌株AZR的基因组中扩增出偶氮还原酶基因,其大小为567 bp,编码188个氨基酸.GenBank搜索表明其为新基因,递交GenBank数据库,获得登录号为EU849488.通过互联网数据库及生物信息学分析工具进行初步分析表明,该基因编码的蛋白属于黄素蛋白家族,是一种依赖NADPH的FMN还原酶;是一种等电点为5.75的稳定蛋白,定位于细胞质中:由α螺旋、延伸带和随机卷曲3种形式组成;具有蛋白激酶C磷酸化等多个位点,并具有一个强跨膜疏水区.%The result of 16S rRNA analysis showed the strain AZR belonged to Staphylococcus cohnii. According to three azoreductase's gene sequences from Staphylococcus in GenBank, a pair of primers were designed and the azoreductase's gene was amplified from the strain AZR's genome successfully. The sequence contained a complete 567 bp ORF (Open Reading Frame) encoding the azoreductase, consisting of 188 amino acids. Blast in GenBank indicated that it is a novel gene, and it has been deposited with the GenBank Data Libraries under accession number EU849488. The characteristics of the protein coded by this gene were analyzed by bioinformatics. The result showed that this protein belonged to the flavin protein family and was a kind of NADPH-depedent FMN reductase, and it was stable, pI 5.75, located in cytoplasm, and composed of Alpha helix, extrended strand random coil. There was a srong transmembrance domain and contained some conserved sites such as protein dinase C-phosphorylation.