Construction and identification of the double auxotrophic Candida parapolymorpha
10.3760/cma.j.issn.0254-5101.2012.11.010
- VernacularTitle:拟多形假丝酵母的双营养缺陷型工程化改造及鉴定
- Author:
Yu LIANG
;
Yuqin JIN
;
Jing ZHANG
;
Shi CHEN
;
Qiming LI
- Publication Type:Journal Article
- Keywords:
Candida parapolymorpha;
Auxotrophic;
ura3 gene;
Leu2 gene
- From:Chinese Journal of Microbiology and Immunology
2012;(11):961-966
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct a genetically-stable double auxotrophic,in which the uracil and leucine were mutated,using the Candida parapolymorpha ATCC26012 as materials.Methods Based on the physical and genetic engineering methods,the chromosome of the C.parapolymorpha strain was modified,where the ura3 and leu2 genes were directly mutated,to obtain the uracil and leucine double auxotrophic strain.Then the constructed strain was identified by the analysis of its biological properties,such as genetic stability,the change of the genes,and the physiologic and biochemical characteristics.Results The uracil and leucine double auxotrophic strain is obtained by screening.The biological identification results show that the obtained strain is genetically stable and the targeted genes are directly altered.In addition,the physiologic and biochemical analyses indicate that the auxotrophic can utilize various kinds of carbon and nitrogen nutrient sources,and its growth is good.Conclusion The successful construction of double auxotrophic mutant strains facilitated the genetic studies on C.parapolymorpha to meet various investigational purposes.Moreover,the constructed auxotrophic strains can be applied as advantageous host cells to express multiple proteins/antigens simultaneously,which is of great significance in the development of vaccines.
