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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。
Objective: To clone Candida albicans hsp60 gene. Methods: The hsp60 gene fragment of Candida albicans was amplified by RT-PCR, purified, ligated with pMD18-T vector, transformed and screened by Amp, x-Gal and IPTG. The white cloning plasmid, SacⅠ and HindⅢ double enzyme Cut and PCR identification, gene sequencing. Results: The 1701bp fragment was amplified by RT-PCR. The recombinant plasmid pMD18-T-hsp60 double digested at 2692bp and 1701bp showed that the sequence of the fragment was similar to the Candida albicans hsp60 gene in GenBank and the homology was 99 %. Conclusion: The successful cloning of Candida albicans hsp60 gene lay the foundation for further study of its immune protection mechanism and preparation of Candida albicans vaccine.