白念珠菌钙调蛋白定点突变重组菌株的构建和鉴定

Construction and identification of calmodulin fixed-point mutant recombinant Candida albican strains

目的 :构建白念珠菌钙调蛋白 (Cm d1)定点突变重组菌株 ,为进一步探讨钙调蛋白基因突变对真菌生长周期及致病性的影响奠定基础。 方法 :将含白念珠菌钙调蛋白基因 (CMD1)定点突变的重组质粒转染 CMD1缺陷 HIS3+重组单倍体菌株 ,用 His/ Trp省却培养基和 His/ Trp/ U ra省却培养基联合选择培养筛选到 Cmd1定点突变酵母菌株。结果 :经选择培养筛选得到能在 His/ Trp省却培养基上生长而不能在 His/ Trp/ Ura省却培养基生长的菌株 4株。结论 :成功构建了 4株 Cm d1定点突变重组菌株 ,为进一步探讨 Cmd1定点突变对白念珠菌生物学特性及致病性的影响奠定基础。

Objective: To construct calmodulin fixed point mutant recombinant Candida albican strains. Methods: The Cmd1 recombinant plasmids were transfected into the monoploid cells of CMD1 defective HIS3 yeast cells respectively. Then the recombinant yeast strains were selected under the selection pressure. Results:We obtained 4 yeast strains which could grow on His/Trp dropout culture medium but couldn't grow on His/Trp/Ura dropout culture medium. Conclusion:Calmodulin fixed point mutant recombinant yeast strains are successfully constructed and will be useful for further research. [