白念珠菌CaPTC4基因的鉴定

发布时间：2018-05-28 00:33

  本文选题：白念珠菌 + CaPTC4　； 参考：《天津大学》2009年硕士论文

【摘要】： 在酿酒酵母中有7种PP2C类蛋白磷酸酯酶,编码它们的基因分别是PTC1、PTC2、PTC3、PTC4、PTC5、PTC6和PTC7。然而,在重要的机会性致病真菌白念珠菌中目前只有CaPTC7已被鉴定,其它的白念珠菌PP2C类蛋白磷酸酯酶尚未见报道。我们将酿酒酵母ScPtc4p的氨基酸序列在白念珠菌数据库中做BLAST(Basic Local Alignment Search Tool)分析,搜寻到酿酒酵母基因ScPTC4的同源序列CaPTC4(orf19.6638)。CaPTC4基因的ORF全长为1038个碱基,编码一个长度为345个氨基酸的蛋白质。白念珠菌CaPtc4p与酿酒酵母ScPtc4p的相同性为35.8%,相似性为48.8%。通过实验我们证明了CaPTC4编码的蛋白质具有磷酸酯酶活性,且活性依赖于镁离子和锰离子并受到抑制剂氟化钠的抑制。这些研究表明CaPTC4编码的蛋白质是一种PP2C类蛋白磷酸酯酶。此外,我们用“URA-blaster”方法对CaPTC4基因进行敲除,得到了白念珠菌CaPTC4基因的完全缺失株(ptc4/ptc4)。构建了表达CaPtc4p绿色荧光融合蛋白的质粒pCR4-CaPTC4GFP,为今后深入研究CaPTC4基因的功能奠定了坚实的基础。
[Abstract]:In Saccharomyces cerevisiae, there are seven kinds of PP2C protein-like phosphatase. The genes encoding them are PTC1, PTC2, PTC3, PTC4, PTC5, PTC6 and PTC7, respectively. However, among the important opportunistic fungi Candida albicans, only CaPTC7 has been identified, and other Candida albicans PP2C protein phosphatase has not been reported. The amino acid sequence of Saccharomyces cerevisiae ScPtc4p was analyzed by BLAST(Basic Local Alignment Search Tool) in Candida albicans database. The homologous CaPTC4(orf19.6638).CaPTC4 gene of Saccharomyces cerevisiae ScPTC4 was found to be 1038 bases in length, encoding a 345-amino acid protein. The similarity between Candida albicans CaPtc4p and Saccharomyces cerevisiae ScPtc4p was 35.8and the similarity was 48.8%. It is proved that the protein encoded by CaPTC4 has phosphatase activity, which is dependent on magnesium and manganese ions and inhibited by sodium fluoride. These studies suggest that the protein CaPTC4 encodes is a PP2C-like protein phosphatase. In addition, we used "URA-blaster" method to knockout the CaPTC4 gene, and obtained the complete deletion strain of CaPTC4 gene of Candida albicans, ptc4 / ptc4. The plasmid pCR4-CaPTC4GFP expressing CaPtc4p green fluorescent fusion protein was constructed, which laid a solid foundation for further study on the function of CaPTC4 gene in the future.
【学位授予单位】：天津大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R379

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