hsp60基因序列分析及其在病原菌鉴定中的应用

目的研究致病菌hsp60基因序列的差异性，为建立致病菌快速鉴定方法及为其他研究提供科学依据。方法在GenBank中查找所需却60基因序列，运用分子生物学软件Primer5．0和Oligo6．0设计合适的通用简并引物，采用合适的扩增条件扩增常见致病菌hsp60基因片段。利用分子生物信息学分析hsp60基因序列的保守性、变异性及菌株间种系进化关系。结果可扩增出常见致病菌hsp60基因片段，序列比较研究显示hsp60基因序列保守与变异并存，保守区与变异区呈“锯齿状”分布。种系进化关系同16SrRNA、23SrRNA分析结果一致。结论hsp60基因序列分布特点为建立致病菌的分子生物学快速鉴别诊断方法如基因芯片研制等提供了可靠的科学依据和重要的实验基础。

Sequence analysis of hsp60 gene and its application in pathogens fast identification

Objective To find, if any, the difference of pathogenic microorganism hsp60 gene and to provide an import,ant basis for fast identification of pathogens. Methods To find out hsp60 gene sequence of pathogens in GenBank and to design proper universal degenerate primer for amplification. With bio-information, the conservative character, the variant character and genetic evolution of bacteria strains were studied. Results hsp60 gene fragment of pathogens can be amplified. The sequences comparative study shows that hsp60 gene is characterized by conservation and variation. Conservative and mutant regions are separately distributed and most small mutant regions distributed in conservative regions. The phylogenetic trees among different bacteria drawn from hsp60 gene were consistent with results from 16S rRNA, 23S rRNA gene. Conclusion The sequence distribution of hsp60 gene would provide a solid basis for the quick identification of pathogens and the construction of a diagnosis mieroarray.