单核细胞增多性李斯特菌新疆分离株lmo0159基因克隆及生物信息学

Cloning and Bioinformatics Analysis of Lmo0159 Gene of Listeria monocytogenes Isolated Strain from Xinjiang

采用PCR方法对lmo0159基因进行扩增、克隆及序列测定,通过应用在线EXPASTYProteomic、SOSUI、SMART、SignalP、TMHMM、SOPMA及ProtFun等程序,同时结合DNAMAN和DNAStar生物信息学软件,对LM90SB2菌株lmo0159基因进行序列分析,并对其编码蛋白质的空间结构和功能预测。结果表明,分离株LM90SB2的lmo0159基因序列全长为2070bp,包含1851bp开放阅读框,共编码617个氨基酸;序列同源比对显示,LM90SB2菌株lmo0159基因核苷酸和氨基酸序列与4b型参考菌株同源性均较高,分别为99.1%~100.0%和99.7%~100.0%。系统进化树显示,LM90SB2菌株lmo0159基因核苷酸序列与LM3、LM9、LM11、LM12、F2365、LM188、NSTN、H34、hs2008、LL195和2306等菌株聚为同一支,说明它们的亲缘关系比较近。lmo0159蛋白质是一种偏酸性、稳定、亲水性蛋白质,其二级结构是混合型,其中无规则卷曲最多,无跨膜区域,无信号肽区域,但含有1个胶原蛋白结合域和3个CnaB结构域。功能分析显示,lmo0159蛋白质在氨基酸生物合成、酶代谢、脂肪酸代谢、免疫应答和胁迫应答等方面的几率均较高。本研究成功克隆LM90SB2lmo0159基因,为进一步研究LM90SB2的lmo0159基因功能提供了帮助。

The lmo0159 gene was amplified, cloned and sequenced by PCR. The sequence of lmo0159 gene of LM90SB2 strain was analyzed, and the spatial structure and function of the lmo0159 protein were predicted by using online softwares, such as EXPASTY Proteomic, SOSUI, SMART, SignalP, TMHMM, SOPMA and ProtFun, also combined with bioinformatics softwares, like DNAMAN and DNAStar. The results showed that the full length of lmo0159 gene sequence of LM90SB2 strain was 2 070 bp with a 1 851 bp open reading frame (ORF), encoding 617 amino acids. Sequence homology comparison showed that the nucleotide and amino acid sequences of lmo0159 gene of LM90SB2 strain had high homology with the strains of 4b serotype, which were 99.1% ~ 100.0% and 99.7%~100.0%, respectively. The phylogenetic tree showed that the nucleotide sequence of lmo0159 gene of LM90SB2 strain was closely relative with LM3, LM9, LM11, LM12, F2365, LM188, NTSN, H34, hs2008, LL195 and 2306 strains, which were in the same branch. The lmo0159 protein was a kind of slightly acidic, stable and hydrophilic protein. It had a mixed-type secondary structure with a large number of random coil. It had no Markiewitransmembrane or signal peptide regions, but a collagen binding domain and 3 Can B (Cna-like, B-type) domains. Function analysis showed that the lmo0159 protein had high odds in amino acid biosynthesis, enzyme metabolism, fatty acid metabolism, immune response and stress response. In this study, the lmo0159 gene of LM90SB2 strain was successfully cloned, which would be helpful to further study the function of lmo0159 gene from LM90SB2 strain.