Cloning, Expression Analysis and Enzyme Activity Assays of the alpha-Carbonic Anhydrase Gene from Chlamydomonas sp ICE-L

doi:10.1007/s12033-017-0040-9

	Cloning, Expression Analysis and Enzyme Activity Assays of the alpha-Carbonic Anhydrase Gene from Chlamydomonas sp ICE-L
	Qu, Changfeng 1,2; He, Yingying 1; Zheng, Zhou 1,2; An, Meiling 1,3; Li, Lulu 1; Wang, Xixi 1; He, Xiaodong 1; Wang, Yibin 1; Liu, Fangming 1; Miao, Jinlai 1,2,3
刊名	MOLECULAR BIOTECHNOLOGY
	2018-01
卷号	60 期号:1 页码:21-30
关键词	Chlamydomonas sp ICE-L alpha-Carbonic anhydrase (alpha-CA) qRT-PCR Bioinformatics analysis Enzyme activity
ISSN号	1073-6085
DOI	10.1007/s12033-017-0040-9
英文摘要	The alpha-carbonic anhydrase (alpha-CA) is a zinc ion-containing enzyme that catalyzes the hydration of carbon dioxide. In this paper, a full-length alpha-CA gene was cloned from Chlamydomonas sp. ICE-L using RT-PCR and RACE-PCR for bioinformatic analysis. The alpha-CA open reading frame obtained by PCR was cloned into a vector and transformed into Escherichia coli to generate alpha-CA-producing bacteria. The alpha-CA was highly expressed upon induction with isopropyl-beta-d-thiogalactoside (IPTG) at a final concentration of 0.8 mM. A single band with a molecular weight of approximate 40 kDa expressed in the recombinant E. coli strain harboring the alpha-CA vector was observed in SDS-PAGE analysis. The carbon dioxide hydration activity and esterase activity of alpha-CA expressed by the recombinant strain were 0.404 U/mg and 0.319 U, respectively. In addition, three conditions, temperature, salinity and UVB radiation exposure, were selected to analyze alpha-CA transcription levels by qRT-PCR. The results suggested UVB exposure increased the expression of relative mRNA; meanwhile, the alpha-CA mRNA expression was rapidly induced by temperature and salinity stress, indicating that Chlamydomonas sp. ICE-L might modulate the alpha-CA mRNA expression to adapt to the extreme environments.
资助项目	Polar Strategic Foundation of China[20150303]
WOS关键词	SOLAR UV-RADIATION ; CAH3 ; PHOTOSYNTHESIS ; TEMPERATURE ; INHIBITION ; EVOLUTION ; MEMBRANES ; BACTERIAL ; GROWTH ; ACID
WOS研究方向	Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology
语种	英语
出版者	HUMANA PRESS INC
WOS记录号	WOS:000419616400003
内容类型	期刊论文
源URL	[http://ir.fio.com.cn:8080/handle/2SI8HI0U/26391]
专题	自然资源部第一海洋研究所
通讯作者	Miao, Jinlai
作者单位	1.State Ocean Adm, Inst Oceanog 1, Key Lab Marine Bioact Subst, Qingdao 266061, Peoples R China 2.Qingdao Natl Lab Marine Sci & Technol, Lab Marine Drugs & Bioprod, Qingdao 266237, Peoples R China 3.Qingdao Univ, Coll Med, Qingdao 266071, Peoples R China
推荐引用方式 GB/T 7714	Qu, Changfeng,He, Yingying,Zheng, Zhou,et al. Cloning, Expression Analysis and Enzyme Activity Assays of the alpha-Carbonic Anhydrase Gene from Chlamydomonas sp ICE-L[J]. MOLECULAR BIOTECHNOLOGY,2018,60(1):21-30.
APA	Qu, Changfeng.,He, Yingying.,Zheng, Zhou.,An, Meiling.,Li, Lulu.,...&Miao, Jinlai.(2018).Cloning, Expression Analysis and Enzyme Activity Assays of the alpha-Carbonic Anhydrase Gene from Chlamydomonas sp ICE-L.MOLECULAR BIOTECHNOLOGY,60(1),21-30.
MLA	Qu, Changfeng,et al."Cloning, Expression Analysis and Enzyme Activity Assays of the alpha-Carbonic Anhydrase Gene from Chlamydomonas sp ICE-L".MOLECULAR BIOTECHNOLOGY 60.1(2018):21-30.