新生隐球菌转录共激活因子MBF1基因的鉴定与敲除  

Identification and knocking-out of transcription co-activation factor gene MBF1 in Cryptococcus neoformans

作　　者：周兆婧 孟云芳[1,2] 赵静宇 桑军军 张超 法振宗 方伟[2] 廖万清[1,2] 

机构地区：[1]上海市医学真菌分子生物学重点实验室,上海200003 [2]上海长征医院皮肤科,上海200003 [3]上海市皮肤病医院,上海200050

出　　处：《中国真菌学杂志》2015年第2期80-83,共4页Chinese Journal of Mycology

基　　金：国家973项目(2013CB531601);上海市自然科学基金(12ZR1454400;12JC1411000);上海市医学真菌分子生物学实验室基金(14DZ2272900)

摘　　要：目的构建新生隐球菌转录共激活因子MBF1基因缺陷菌株。方法采用套叠PCR方法 ,构建含有抗性基因NEO以及靶基因上下游同源DNA片段的基因敲除框,通过基因枪将重组片段转化入新生隐球菌,应用PCR筛选和DNA序列测序方法对基因突变株进行鉴定与验证。结果成功构建了新生隐球菌基因突变株mbf1裣。结论通过基因突变株mbf1裣的构建,为深入研究新生隐球菌转录辅助因子Mbf1的功能机制奠定基础。Objective To construct the mutant strain of transcriptional coactivated factor Mbf1 in Cryptococcus neoformans . Methods Amplified the homologous recombination cassette containing report gene NEO and flanking DNA fragments of MBF1 via overlap PCR,which was then transformed into Cryptococcus neoformans through gene gun.The positive transformants was screened by diagnostic PCR and then confirmed by DNA sequencing.Results The target gene MBF1 was successfully knocked out in Cryptococcus neoformans.Conclusion Construction of the mutant strain would lay the foundation for further pathogenetic studies research of transcriptional coactivated factor Mbf1 in Cryptococcus neoformans.

关 键 词：新生隐球菌 致病机制 基因敲除 

分 类 号：R379.5[医药卫生—病原生物学]