吕倩,王颜颜,刘涛华,牟丽丽,陈燕,康颖倩
LV Qian,WANG Yanyan,LIU Taohua,MOU Lili,CHEN Yan,KANG Yingqian

• 1:贵州医科大学微生物学教研室

摘要(Abstract)：

目的:优化念珠菌RNA聚合酶Ⅱ大亚基(RPB1)基因片段的PCR扩增条件,筛选适宜念珠菌RPB1的PCR扩增体系。方法:使用改良CTAB法提取念珠菌基因组DNA,分别调整念珠菌RPB1基因片段PCR扩增过程中dd H2O含量、DNA模板含量、退火温度或循环次数,比较不同反应体系的PCR扩增效果。结果:在26μL的PCR反应体系中,DNA模板含量2μL、退火温度为56℃、循环40次时是念珠菌RPB1基因片段最稳定的PCR扩增反应体系,扩增得到24条皱褶念珠菌RPB1基因片段序列,上传至Gen Bank。结论:优化PCR扩增条件后,念珠菌RPB1基因片段的目的条带更明亮、清晰,为基因测序及深入研究念珠菌的工作奠定了基础。
Objective: To optimize PCR amplification condition of Candida RNA polymerase Ⅱ rbcl(RPB1) gene fragment,screening appropriate PCR amplification system of Candida RPB1. Methods:Adopting modified CTAB method to extract Candida genome DNA,adjusting dd H2 O content of Candida RPB1 gene fragment PCR amplification process,DNA mould content,annealing or number of cycles respectively; comparing PCR amplification effect of different response system. Results: In 26 μL PCR response system,Candida RPB1 gene fragment was in the most stable PCR amplification amplification when DNA mould content was 2 μL,annealing was 56 ℃ and number of cycles was 40; amplification yielded 24 strains of Candida rugosa RPB1 gene fragment sequence and uploaded to Gen Bank.Conclusion: After optimizing PCR amplification conditions,target band of Candida RPB1 gene fragment was obvious and clear,which laid the foundation of gene sequencing and in-depth research of Candida.

关键词(KeyWords)： 念珠菌属;RPB1基因片段;CTAB;基因扩增
Candida;RPB1 gene fragment;CTAB;gene amplification

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金(31060006;31260029);; 贵州省社会发展科技攻关项目[黔科合SY字(2011)3017号];; 贵阳市科技局社会发展与民生计划[筑科合同(201103)16号]

作者(Author): 吕倩,王颜颜,刘涛华,牟丽丽,陈燕,康颖倩
LV Qian,WANG Yanyan,LIU Taohua,MOU Lili,CHEN Yan,KANG Yingqian

DOI: 10.19367/j.cnki.1000-2707.2016.02.004

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