Gene cloning, sequence analysis, and expression profiles of a novel beta-ring carotenoid hydroxylase gene from the photoheterotrophic green alga Chlorella kessleri

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	Gene cloning, sequence analysis, and expression profiles of a novel beta-ring carotenoid hydroxylase gene from the photoheterotrophic green alga Chlorella kessleri
	Yu, Xiaona 1; Cui, Hongli 2,3; Cui, Yulin2 ; Wang, Yan2 ; Li, Xueqin 4; Liu, Zhaopu 1; Qin, Song1,2
刊名	MOLECULAR BIOLOGY REPORTS
	2014-11-01
卷号	41 期号:11 页码:7103-7113
关键词	Carotenoid hydroxylase Chlorella kessleri Gene cloning High light Photoheterotrophic
ISSN号	0301-4851
通讯作者	Liu, ZP (reprint author), Nanjing Agr Univ, Key Lab Marine Biol, Coll Resources & Environm Sci, Nanjing 210095, Jiangsu, Peoples R China. sea@njau.edu.cn ; sqin@yic.ac.cn
产权排序	[Yu, Xiaona; Liu, Zhaopu; Qin, Song] Nanjing Agr Univ, Key Lab Marine Biol, Coll Resources & Environm Sci, Nanjing 210095, Jiangsu, Peoples R China; [Cui, Hongli; Cui, Yulin; Wang, Yan; Qin, Song] Chinese Acad Sci, Yantai Inst Coastal Zone Res, Key Lab Coastal Biol & Biol Resources Utilizat, Yantai 264003, Peoples R China; [Cui, Hongli] Univ Chinese Acad Sci, Beijing 100049, Peoples R China; [Li, Xueqin] Shenzhen Univ, Coll Life Sci, Shenzhen Key Lab Marine Bioresource & Ecoenvironm, Shenzhen 518060, Peoples R China
中文摘要	In this study, a full-length complementary DNA (cDNA) sequence of beta-ring carotenoid hydroxylase (CHY), designated Ckecyp97a1, was isolated via reverse transcription-polymerase chain reaction and rapid amplification of cDNA ends (RACE) methods. The cloned Ckecyp97a1 cDNA was 2,264-bp in length, and contained an open reading frame (ORF) of 1,944-bp with 5'-terminal untranslated region (UTR) of 66-bp and 3'-terminal UTR of 254-bp and encoded a beta-ring CHY protein of 647 amino acids. The deduced protein had a calculated molecular mass of 71.43 kDa with an estimated isoelectric point (pI) of 6.72. Multiple sequence alignment and phylogenetic analysis revealed that Ckecyp97a1 was homologs to known chloroplastic cytochrome P450 (P450) CHY. The typical catalytic motifs of the P450 were highly conserved in the protein sequences of CkeCYP97A1. The Ckecyp97a1 transcriptional expression and carotenoids accumulation were observed under high light (HL) of different wavelengths (white: 390-770 nm and blue: 420-500 nm). The results revealed that Ckecyp97a1 transcript increased strongly throughout the course of the HL illumination treatment (22-70 h) under white HL treatment, while decreased during 10-58 h under blue HL treatment. The concentrations of lutein, alpha-carotene, and beta-carotene were relatively steady and below the control level under both treatments. The zeaxanthin concentration was higher under white HL treatment than those under control and blue HL treatments. Ckecyp97a1 gene showed different expression patterns under different light wavelengths treatments. The data obtained in this study demonstrates that CkeCYP97A1 is the enzyme responsible for carotenoid hydroxylation involved in HL acclimation for photoheterotrophic green alga Chlorella kessleri CGMCC 4917.
英文摘要	In this study, a full-length complementary DNA (cDNA) sequence of beta-ring carotenoid hydroxylase (CHY), designated Ckecyp97a1, was isolated via reverse transcription-polymerase chain reaction and rapid amplification of cDNA ends (RACE) methods. The cloned Ckecyp97a1 cDNA was 2,264-bp in length, and contained an open reading frame (ORF) of 1,944-bp with 5'-terminal untranslated region (UTR) of 66-bp and 3'-terminal UTR of 254-bp and encoded a beta-ring CHY protein of 647 amino acids. The deduced protein had a calculated molecular mass of 71.43 kDa with an estimated isoelectric point (pI) of 6.72. Multiple sequence alignment and phylogenetic analysis revealed that Ckecyp97a1 was homologs to known chloroplastic cytochrome P450 (P450) CHY. The typical catalytic motifs of the P450 were highly conserved in the protein sequences of CkeCYP97A1. The Ckecyp97a1 transcriptional expression and carotenoids accumulation were observed under high light (HL) of different wavelengths (white: 390-770 nm and blue: 420-500 nm). The results revealed that Ckecyp97a1 transcript increased strongly throughout the course of the HL illumination treatment (22-70 h) under white HL treatment, while decreased during 10-58 h under blue HL treatment. The concentrations of lutein, alpha-carotene, and beta-carotene were relatively steady and below the control level under both treatments. The zeaxanthin concentration was higher under white HL treatment than those under control and blue HL treatments. Ckecyp97a1 gene showed different expression patterns under different light wavelengths treatments. The data obtained in this study demonstrates that CkeCYP97A1 is the enzyme responsible for carotenoid hydroxylation involved in HL acclimation for photoheterotrophic green alga Chlorella kessleri CGMCC 4917.
学科主题	Biochemistry & Molecular Biology
研究领域[WOS]	Biochemistry & Molecular Biology
关键词[WOS]	HAEMATOCOCCUS-PLUVIALIS ; ASTAXANTHIN PRODUCTION ; FUNCTIONAL-ANALYSIS ; BIOSYNTHESIS GENES ; IN-VIVO ; ARABIDOPSIS ; LIGHT ; ACCUMULATION ; PLANTS ; PHOTOPROTECTION
收录类别	SCI
语种	英语
WOS记录号	WOS:000344101300008
内容类型	期刊论文
源URL	[http://ir.yic.ac.cn/handle/133337/8526]
专题	烟台海岸带研究所_海岸带生物学与生物资源利用所重点实验室
作者单位	1.Nanjing Agr Univ, Key Lab Marine Biol, Coll Resources & Environm Sci, Nanjing 210095, Jiangsu, Peoples R China 2.Chinese Acad Sci, Yantai Inst Coastal Zone Res, Key Lab Coastal Biol & Biol Resources Utilizat, Yantai 264003, Peoples R China 3.Univ Chinese Acad Sci, Beijing 100049, Peoples R China 4.Shenzhen Univ, Coll Life Sci, Shenzhen Key Lab Marine Bioresource & Ecoenvironm, Shenzhen 518060, Peoples R China
推荐引用方式 GB/T 7714	Yu, Xiaona,Cui, Hongli,Cui, Yulin,et al. Gene cloning, sequence analysis, and expression profiles of a novel beta-ring carotenoid hydroxylase gene from the photoheterotrophic green alga Chlorella kessleri[J]. MOLECULAR BIOLOGY REPORTS,2014,41(11):7103-7113.
APA	Yu, Xiaona.,Cui, Hongli.,Cui, Yulin.,Wang, Yan.,Li, Xueqin.,...&Qin, Song.(2014).Gene cloning, sequence analysis, and expression profiles of a novel beta-ring carotenoid hydroxylase gene from the photoheterotrophic green alga Chlorella kessleri.MOLECULAR BIOLOGY REPORTS,41(11),7103-7113.
MLA	Yu, Xiaona,et al."Gene cloning, sequence analysis, and expression profiles of a novel beta-ring carotenoid hydroxylase gene from the photoheterotrophic green alga Chlorella kessleri".MOLECULAR BIOLOGY REPORTS 41.11(2014):7103-7113.