沙门菌grOEL基因序列在系统发育分析和PCR-限制性片段多态性分析鉴定中的应用

Phylogenetic analysis and PCR-restriction fragment length polymophism identification of Salmonella based on grOEL gene sequence

目的探讨grOEL基因序列在沙门菌进化分析和分型鉴定中的应用。方法对8种血清群的沙门菌菌株进行PCR扩增、测序,然后用分子生物学软件Bioedit与DNAstar进行序列分析,同时用PCR-限制性片段多态性分析(RFLP)技术进行分型鉴定。结果8种沙门菌血清群grOEL基因保守序列与变异序列呈锯齿状排列,其grOEL氨基酸系统发育树与grOEL基因系统发育树结果不完全相同,但O8、O9、O10之间亲缘关系最近,PCR-RFLP分析有5种模式图。结论grOEL基因序列在沙门菌系统发育中可作为遗传标记,同时也可作为分型鉴定的靶序列。

Objective To investigate the value of grOEL gene sequence in phylogenetic analysis and typing of Salmonella. Methods The grOEL gene was amplified by PCR, sequenced and analyzed using Bioedit and DNAstar software. The Salmonella strains were identified using PCR-restriction fragment length polymophism （PCR-RFLP）. Results The conservative and variable regions ofgrOEL gene of Salmonella serogroup were separately distributed and most of the small mutant regions distributed intermittently among the conservative regions. The phylogenetic tree of Salmonella based on the nucleotides differed from that generated based on the amino acid sequence. O8, O9 and O10 had the closest consanguinity, and 5 patterns were identified by PCR-RFLP. Conclusion The grOEL gene can be used as a genetic marker for phylogenetic analysis of Salmonella and also as a target sequence for Salmonella typing identification.