本文選題：白念珠菌 + CaRCH1��； 參考：《天津大學(xué)》2010年碩士論文

【摘要】：白念珠菌RCH1基因的ORF.長為1236 bp,編碼一個411個氨基酸的蛋白,由于該基因的缺失株對氟康唑有抗性表型,所以曾經(jīng)命名為CaFSP1。在課題研究中,我們發(fā)現(xiàn)rch1Δ/rch1Δ缺失株和pmc1Δ/pmc1Δ缺失株具有類似的表型,都對鈣離子敏感,對LiCl和氟康唑有抗性的表型。我們知道環(huán)孢霉素A(CsA)可以抑制鈣調(diào)磷酸酶,阻斷白念珠菌Ca~(2+)/鈣調(diào)磷酸酶途徑。于是,首先我們將CsA加入CaCl_2的固體培養(yǎng)基中,rch1Δ/rch1Δ缺失株可以回復(fù)到野生型的生長狀態(tài)。第二,胞外低濃度的Mg~(2+)能夠抑制pmc1Δ/pmc1Δ缺失株對鈣離子的敏感表型,我們將Mg~(2+)加入CaCl_2培養(yǎng)基中,rch1Δ/rch1Δ缺失株對CaCl2敏感的表型消失。第三,我們構(gòu)建了pmc1Δ/pmc1Δrch1Δ/rch1Δ雙基因缺失株,發(fā)現(xiàn)該缺失株與pmc1Δ/pmc1Δ缺失株相比,對鈣離子更加敏感,而對LiCl和氟康唑的表型沒有明顯的差異。此外,鈣調(diào)磷酸酶影響白念珠菌菌落的形態(tài)和菌絲的形成,我們發(fā)現(xiàn)pmc1Δ/pmc1Δrch1Δ/rch1Δ雙基因缺失株在血清誘導(dǎo)下,菌絲的形成受到抑制。然而,在體內(nèi)實驗中,我們發(fā)現(xiàn)白念珠菌僅缺失PMC1基因,就會大大減弱細胞的毒力。最后,我們又構(gòu)建了pCK-RCH1質(zhì)粒,該質(zhì)�？梢赃^量表達RCH1基因,將其轉(zhuǎn)入pmc1Δ/pmc1Δ缺失株中,但未能彌補pmc1Δ/pmc1Δ缺失株的表型。以上實驗可以說明CaRch1p具有調(diào)節(jié)細胞內(nèi)鈣離子穩(wěn)態(tài)的作用,因此我們將該基因重新命名為CaRCH1即C. albicans regulator of calcium homeostasis。
[Abstract]:ORF of RCH1 gene of Candida albicans. The protein, which is 1236 BP long and encodes a 411-amino acid protein, was once named CaFSP1 because of its resistant phenotype to fluconazole. In our study, we found that rch1 螖 / rch1 螖 deletion strain and pmc1 螖 / pmc1 螖 deletion strain had similar phenotypes, which were sensitive to calcium ions and resistant to LiCl and fluconazole. It is known that cyclosporin A can inhibit calmodulase and block the Ca~(2 / Ca 2 + pathway of Candida albicans. So, first, we added CsA to CaCl_2 solid medium and the missing strain of rch1 螖 / rch1 螖 could return to the growth state of wild type. Secondly, the low concentration of Mg~(2 could inhibit the sensitive phenotype of pmc1 螖 / pmc1 螖 deletion strain to Ca ~ (2 +). Mg~(2) was added to CaCl_2 medium to eliminate the CaCl2 sensitive phenotype of rch1 螖 / rch1 螖 deletion strain. Thirdly, we constructed pmc1 螖 / pmc1 螖 rch1 螖 / rch1 螖 double gene deletion strain. It was found that the deletion strain was more sensitive to calcium than that of pmc1 螖 / pmc1 螖 deletion strain, but had no obvious difference in phenotype of LiCl and fluconazole. In addition, calmodulase affected the morphology and mycelium formation of Candida albicans colony. We found that the formation of hyphae was inhibited by pmc1 螖 / pmc1 螖 rch1 螖 / rch1 螖 double gene deletion strain induced by serum. However, in vivo experiments, we found that Candida albicans can significantly reduce the virulence of cells by simply missing the PMC1 gene. Finally, we constructed the pCK-RCH1 plasmid, which can overexpress the RCH1 gene and transfer it into the pmc1 螖 / pmc1 螖 deletion strain, but it can not make up for the phenotype of the pmc1 螖 / pmc1 螖 deletion strain. These results suggest that CaRch1p can regulate intracellular calcium homeostasis, so we rename the gene CaRCH1 or C. albicans regulator of calcium homeostasis..
【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】：2010
【分類號】：R379

【引證文獻】

相關(guān)博士學(xué)位論文 前1條

1 劉偉;白念珠菌CaSCH9基因功能研究[D];天津大學(xué);2010年

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本文編號：1873635