[新型隐球菌cap70转化系统的构建]
[Construction of Cryptococcus neoformans cap70 transformation system].
作者信息
Pan W, Liao W, Huo K
机构信息
Department of Dermatology, Changzheng Hospital, The Second military Medical University, Shanghai 200003, China.
出版信息
Zhonghua Yi Xue Za Zhi. 2001 Jun 25;81(12):748-51.
OBJECTIVE
To construct a transformation system of Cryptococcus neoformans capsule-deficient strain cap70.
METHODS
(1)Obtained ura5 mutants by screening the Cryptococcus neoformans cap70 using 5-fluoroorotic acid counter selection method, analyzed the obtaining strain by Southern blot and PCR; (2)Through the plasmid pCXJ18 and pCXJU which contained ura5 gene from Cryptococcus neoformans and ura3 from Kluyveromyces fragilils. as a selective marker, the ura5 strains were transformed by electrotransformation and chemical transformation.
RESULTS
Obtained ura5 mutants, they were detected as lack of ura function. The chemical transformation method was first be used in Cryptococcus neoformans transformation.
CONCLUSION
A transformation system of Cryptococcus neoformans cap70 has been established, which provides not only the basis for gene cloning from the species for studying gene function and gene expression.
目的
构建新型隐球菌荚膜缺陷型菌株cap70的转化系统。
方法
（1）采用5-氟乳清酸反向选择法筛选新型隐球菌cap70，获得ura5突变体，通过Southern杂交和PCR对获得的菌株进行分析；（2）以含有新型隐球菌ura5基因和脆壁克鲁维酵母ura3基因的质粒pCXJ18和pCXJU作为选择标记，通过电转化和化学转化对ura5菌株进行转化。
结果
获得ura5突变体，检测发现其缺乏ura功能。化学转化方法首次应用于新型隐球菌的转化。
结论
已建立新型隐球菌cap70的转化系统，这不仅为从该物种克隆基因以研究基因功能和基因表达提供了基础。
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