白色念珠菌hsp60基因克隆

目的:克隆白色念珠菌hsp60基因。方法:通过RT-PCR技术扩增白色念珠菌hsp60基因片段,纯化后与pMD18-T载体连接,转化并通过Amp、x-Gal、IPTG进行蓝白筛选,白色克隆提取质粒,SacⅠ和HindⅢ双酶切及PCR鉴定,基因测序。结果:RT-PCR扩增得到1701bp片断,重组质粒pMD18-T-hsp60双酶切后在2692bp和1701bp处可见到酶切片段,测序结果与GenBank中白色念珠菌hsp60基因比较,同原性达99%。结论:成功克隆白色念珠菌hsp60基因,为进一步研究其免疫保护机制及制备白色念珠菌疫苗奠定基础。

Clone of heat shock protein 60 gene of candida albicans

Objective:To clone the heat shock protein 60 (hsp60) gene of candida albicans.Methods:Hsp60 gene fragments of candida albicans were amplified by RT-PCR, then purified gene fragments were inserted into pMD18-T expression vector, the clones were screened by white-blue screening,plasmids were extracted from white clone, genetic sequencing was conducted.Results:A 1 701 bp fragment was obtained by RT-PCR amplification in the plasmid of pMD18-hsp60 and the homogeneity was up to 99% compared with hsp60 gene sequence of candida albicans.Conclusion:Cloning the hsp60 gene of candida albicans successfully provides a basis for immunoprotection mechanism investigation and anti-candida vaccine preparation.