Construction of Candida albicans two-hybrid library and screening for proteins interacting with Crk1

	Construction of Candida albicans two-hybrid library and screening for proteins interacting with Crk1
	Ni, J; Chen, X; Yang, T; Chen, JY
刊名	ACTA BIOCHIMICA ET BIOPHYSICA SINICA
	2001
卷号	33 期号:2 页码:198-204
关键词	Candida albicans CRK1 LexA two-hybrid system
通讯作者	,
英文摘要	In order to study interactions among proteins involved in Candida albicans morphorgenesis, C. albicans genomic DNA was digested with Sau3AI and fused to activating domain(AD) of LEXA to construct an AD-fused C. albicans genomic DNA expression library. The library contained 7.7 x 10(4) independent: clones and included 1.2 x 10(5) kb DNA which was 10 fold of the C. albicans whole genomic DNA. The CRK1 gene encodes a CDC2-related protein, that was involved in morphorgenesis of C. albicans. The Crk1 fused to DNA binding domain of LexA was used as the bait to screen the LexA library. Among 9 x 10(5) transformants, eight Leu and LacZ double positive clones were selected. With restriction enzyme analysis, three kinds of fragments were identified. The clones NJ1, NJ2, NJ3 were sequenced and analyzed with BLAST program. It was found that NJ1, NJ2, NJ3 were homologs of S. cerevisiae gene STI1, RET2 and NFI1, respectively. The kinase domain of Crk1(Crk1N) interacted weakly with protein encoded by NJ1, while the noncatalytic domain of Crk1 (Crk1C) interacted strongly with proteins encoded by NJ2 and NJ3. The interaction of NJ3 and Crk1C was verified by the coimmunoprecipition of NJ3-HA with Crk1C-LexA. These proteins may be involved in maturation, transport, localization, and activity regulation of the Crk1.
学科主题	Biochemistry & Molecular Biology; Biophysics
类目[WOS]	Biochemistry & Molecular Biology ; Biophysics
关键词[WOS]	GENE
收录类别	SCI
语种	中文
WOS记录号	WOS:000167681800009
内容类型	期刊论文
版本	出版稿
源URL	[http://202.127.25.143/handle/331003/2610]
专题	上海生化细胞研究所_上海生科院生化细胞研究所
推荐引用方式 GB/T 7714	Ni, J,Chen, X,Yang, T,et al. Construction of Candida albicans two-hybrid library and screening for proteins interacting with Crk1[J]. ACTA BIOCHIMICA ET BIOPHYSICA SINICA,2001,33(2):198-204.
APA	Ni, J,Chen, X,Yang, T,&Chen, JY.(2001).Construction of Candida albicans two-hybrid library and screening for proteins interacting with Crk1.ACTA BIOCHIMICA ET BIOPHYSICA SINICA,33(2),198-204.
MLA	Ni, J,et al."Construction of Candida albicans two-hybrid library and screening for proteins interacting with Crk1".ACTA BIOCHIMICA ET BIOPHYSICA SINICA 33.2(2001):198-204.