沼泽红假单胞菌偶氮还原酶相关基因的克隆及其生物信息学分析

Cloning and Bioinformatic Analysis of a Relative Azoredutase Gene from Rhodopsedomonas palustris

脱色实验证明沼泽红假单胞菌 (Rhodopsedomonaspalustris)对偶氮染料有较强的降解能力 ,作者通过GenBank搜索 ,对所获得的所有偶氮还原酶基因在NCBI进行比对并设计引物 ,从沼泽红假单胞菌质粒中扩增获得了一条含 4 71bp完整开放阅读框架的序列。GenBank搜索表明该基因为未登录的新基因。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 :该基因编码的蛋白是一种等电点为 9.6 5的不稳定蛋白 ,定位于细胞质 ;由α螺旋、延伸带和随机卷曲三种形式组成 ,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。

Rhodopsedomonas palustris could degrade in 24h over 75% azo dyes when the original concentration of azo dyes was 100mg/L.Nucleic acid sequences of azoreductase were searched and blasted in GenBank.A pair of primers based on the conserved regions were designed. A specific fragment was amplified by PCR from the plasmid of Rhodopsedomonas palustris and sequenced.The sequence contained a complete 471bp ORF(Open Reading Frame).BLAST in GenBank indicated that it is novel gene.The characteristics of the protein coded by this gene were analyzed by bioinformatics.The result showed that this protein was instable,pI 9.65,located in cytoplasm,and composed of Alpha helix,extrended strand and random coil.There was a srong transmembrance domain and contained some conserved sites such as protein dinase C phosphorylation.