本文選題：白念珠菌 + CaPTC4��； 參考：《天津大學(xué)》2009年碩士論文

【摘要】： 在釀酒酵母中有7種PP2C類蛋白磷酸酯酶,編碼它們的基因分別是PTC1、PTC2、PTC3、PTC4、PTC5、PTC6和PTC7。然而,在重要的機會性致病真菌白念珠菌中目前只有CaPTC7已被鑒定,其它的白念珠菌PP2C類蛋白磷酸酯酶尚未見報道。我們將釀酒酵母ScPtc4p的氨基酸序列在白念珠菌數(shù)據(jù)庫中做BLAST(Basic Local Alignment Search Tool)分析,搜尋到釀酒酵母基因ScPTC4的同源序列CaPTC4(orf19.6638)。CaPTC4基因的ORF全長為1038個堿基,編碼一個長度為345個氨基酸的蛋白質(zhì)。白念珠菌CaPtc4p與釀酒酵母ScPtc4p的相同性為35.8%,相似性為48.8%。通過實驗我們證明了CaPTC4編碼的蛋白質(zhì)具有磷酸酯酶活性,且活性依賴于鎂離子和錳離子并受到抑制劑氟化鈉的抑制。這些研究表明CaPTC4編碼的蛋白質(zhì)是一種PP2C類蛋白磷酸酯酶。此外,我們用“URA-blaster”方法對CaPTC4基因進行敲除,得到了白念珠菌CaPTC4基因的完全缺失株(ptc4/ptc4)。構(gòu)建了表達CaPtc4p綠色熒光融合蛋白的質(zhì)粒pCR4-CaPTC4GFP,為今后深入研究CaPTC4基因的功能奠定了堅實的基礎(chǔ)。
[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.
【學(xué)位授予單位】：天津大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2009
【分類號】：R379

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本文編號：1944478