Cloning and transcription analysis of the Candida rugosa propionyl-CoA dehydrogenase gene and its expression in Pichia pastoris

doi:10.1002/jobm.201100050

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	Cloning and transcription analysis of the Candida rugosa propionyl-CoA dehydrogenase gene and its expression in Pichia pastoris
	Zhou, Feng-Li; Zhang, Ru-Bing; Zhang, Yong-Guang; Zhang, Fan; Zhu, Jian-Hang
	2012-06-01
发表期刊	JOURNAL OF BASIC MICROBIOLOGY
卷号	52 期号:3 页码:360-367
摘要	The propionyl-CoA dehydrogenase (PACD) gene was cloned from Candida rugosa by the cDNA RACE technique. The full cDNA of the PACD gene has a length of 1408 bp, which contains a complete open reading frame (ORF) of 1329 bp, coding for 442 amino acids. The cDNA of PACD was cloned into the expression plasmid pPIC9K and transformed into Pichia pastoris GS115. The recombinant protein was purified by Ni-NTA affinity chromatography, and its size was observed to be approximately 49 kDa as estimated by SDS-PAGE. Anti-His antibodies were used to characterise the recombinant PACD by western-blot analysis. The recombinant protein retained the activity of catalysing propionyl-CoA to acryloyl-CoA. The results of dot-blotting hybridisation using a PACD cDNA probe indicated that the PACD mRNA level was modified at different stages: mRNA levels were low for the first 36 h, then increased through 48 h and eventually reached a stable level. These results indicate that propionate induction could significantly activate PACD mRNA expression. Information from this study will be helpful in elucidating the metabolic pathway for 3-hydroxypropionic acid production in C. rugosa. ((c) 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)
文章类型	Article
关键词	Cloning Expression Transcription Analysis Propionyl-coa Dehydrogenase
WOS标题词	Science & Technology ; Life Sciences & Biomedicine
DOI	10.1002/jobm.201100050
关键词[WOS]	BETA-OXIDATION ; PATHWAY ; ACID ; CATABOLISM ; COENZYME ; COMPLEX
收录类别	SCI
语种	英语
WOS研究方向	Microbiology
WOS类目	Microbiology
WOS记录号	WOS:000305064700015
引用统计	被引频次：2[WOS] [WOS记录] [WOS相关记录]
文献类型	期刊论文
条目标识符	http://ir.qibebt.ac.cn/handle/337004/6072
专题	生物基材料组群
作者单位	Chinese Acad Sci, Qingdao Inst Bioenergy & Bioproc Technol, Key Lab Biofuel, Qingdao 266101, Peoples R China
推荐引用方式 GB/T 7714	Zhou, Feng-Li,Zhang, Ru-Bing,Zhang, Yong-Guang,et al. Cloning and transcription analysis of the Candida rugosa propionyl-CoA dehydrogenase gene and its expression in Pichia pastoris[J]. JOURNAL OF BASIC MICROBIOLOGY,2012,52(3):360-367.
APA	Zhou, Feng-Li,Zhang, Ru-Bing,Zhang, Yong-Guang,Zhang, Fan,&Zhu, Jian-Hang.(2012).Cloning and transcription analysis of the Candida rugosa propionyl-CoA dehydrogenase gene and its expression in Pichia pastoris.JOURNAL OF BASIC MICROBIOLOGY,52(3),360-367.
MLA	Zhou, Feng-Li,et al."Cloning and transcription analysis of the Candida rugosa propionyl-CoA dehydrogenase gene and its expression in Pichia pastoris".JOURNAL OF BASIC MICROBIOLOGY 52.3(2012):360-367.