中华皮肤科杂志 ›› 1999, Vol. 32 ›› Issue (5): 300-302.

• 论著 • 上一篇    下一篇

新生隐球菌cap60荚膜缺陷株转化系统的构建

王珏1, 霍克克2, 廖万清1   

  1. 1. 上海第二军医大学附属长征医院皮肤科 200003;
    2. 复旦大学遗传所遗传工程国家重点实验室
  • 收稿日期:1998-10-13 修回日期:1999-04-06 出版日期:1999-10-15 发布日期:1999-10-15
  • 基金资助:
    国家自然科学基金!(39570043);863计划!(102-11-02-02)资助

Construction of the Cryptococcus Neoformans cap60 Transformation System

WANG Jue1, HUO Keke2, LIAO Wanqing1   

  1. Department of Dermatology, No.6 Hospital, Shanghai 200233
  • Received:1998-10-13 Revised:1999-04-06 Online:1999-10-15 Published:1999-10-15

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摘要： 目的 构建新生隐球菌荚膜缺陷株cap60的转化系统.方法与结果 ①在新生隐球菌的荚膜缺陷株cap60中利用 5-氟乳清酸 (5-FOA)反选择法筛选到一个尿嘧啶合成基因突变株 (cap60 ura5);②利用带有以新生隐球菌ura5基因为选择标记的质粒载体 pCntel1-d通过电转化法和化学转化法成功地转化该ura5突变株;③经质粒稳定性检验和Southern杂交分析表明质粒载体pCntel1-d在新生隐球菌cap60ura5菌株中能以整合和游离两种状态存在.结论首次建立了新生隐球菌cap60荚膜缺陷株的转化系统,这不仅为克隆与荚膜形成有关的新基因提供了基础,还为以基因中断、基因替代方法研究基因功能及基因表达等方面的工作创造了条件.

关键词: 隐球菌,新型, 转化

Abstract: Objective To construct a transformation system of Cryptococcus neoformans.(C-neoformans) capsule deficient strain cap60.Methods and Results The results included:(1) A ura 5 mutant was obtained by screening the C-neoformans cap60 using 5 fluoroorotic acid counter selection method.(2) By electrotransformation and chemical transformation,the ura5 strain was successfully transformed through the plasmid pCntel1-d which contained the ura5 gene from C-neoformans as a selective marker.(3) The plasmid stability test and Southern blot analysis indicated there were two status of plasmid pCntel1-d in C-neoformans transformants:integrated or episomal plasmid.Conclusion A transformation system of C-neoformans cap60 has been established,which provides not only the basis for gene cloning from the species but also for studying gene function and gene expression.

Key words: Cryptococcus neoforman, Transformation