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Molecular cloning and expression analysis of RbcL cDNA from the bloom-forming green alga Chaetomorpha valida (Cladophorales, Chlorophyta)
Deng, Yunyan1; Zhan, Zifeng1; Tang, Xiaorong2; Ding, Lanping3; Duan, Delin1; Duan, DL (reprint author), Chinese Acad Sci, Inst Oceanol, Qingdao 266071, Peoples R China.
2014-08-01
发表期刊JOURNAL OF APPLIED PHYCOLOGY
卷号26期号:4页码:1853-1861
文章类型Article
摘要Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) is a crucial enzyme catalyzing the CO2-fixing reaction in the initial step of the Calvin cycle. In this study, the full-length cDNA of Rubisco large subunit (RbcL) from the bloom-forming green alga Chaetomorpha valida (designated as CvRbcL) was isolated based on homologous cloning and the rapid amplification of cDNA ends (RACE) (accession: KF514135). It was 1,966-bp long, with an open reading frame of 1,431 bp without intron. The deduced 476 amino acids gave a predicted molecular weight of 52.38 kDa and theoretical isoelectric point of 6.15. Three catalytic regions and a CO2 activator region were identified in the CvRbcL, which highly conserved with those of other green algae and higher plants. Homologous analysis indicated that CvRbcL shared 52-86 % similarities with other 13 known algal and higher plant RbcLs sequences. Phylogenetic analysis showed that it had closer relationship with those of green algae and higher plants but was comparatively far from the red and brown algae clade. Real-time PCR detection revealed that the CvRbcL expression were dramatically upregulated by light and strongly suppressed in the dark. Temperature also markedly affected the CvRbcL transcription. A relatively high transcript level appeared between 15-25 A degrees C and reached the maximum at 20 A degrees C. This indirectly reflected that highly expressed Rubisco was involved with CO2 metabolism of C. valida under favorable natural environments, and might contribute to its fast growth and bloom formation. In vitro expression of CvRbcL showed that one distinct band existed at similar to 55 kDa, and western blot detection proved that it was positive to the anti-His antibody with high specificity. Our study is the first to characterize RbcL gene from Chaetomorpha species providing more knowledge of RbcL properties and facilitating further exploration of Rubisco in green macroalgae. Our results also help to decipher Rubisco molecular functions in the rapid growth period of C. valida and provide clues to develop forecasts of the seasonal C. valida bloom.
关键词Bloomgreen Alga Chaetomorpha Valida Green Tide Ribulose-1 5-bisphosphate Carboxylase/oxygenase Large Subunit
学科领域Biotechnology & Applied Microbiology ; Marine & Freshwater Biology
DOI10.1007/s10811-013-0208-z
收录类别SCI
语种英语
WOS记录号WOS:000340101100024
引用统计
被引频次:13[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.qdio.ac.cn/handle/337002/24130
专题实验海洋生物学重点实验室
通讯作者Duan, DL (reprint author), Chinese Acad Sci, Inst Oceanol, Qingdao 266071, Peoples R China.
作者单位1.Chinese Acad Sci, Inst Oceanol, Qingdao 266071, Peoples R China
2.Ocean Univ China, Minist Educ, Key Lab Marine Genet & Breeding, Qingdao 266003, Peoples R China
3.Shantou Univ, Shantou 515063, Peoples R China
第一作者单位中国科学院海洋研究所
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GB/T 7714
Deng, Yunyan,Zhan, Zifeng,Tang, Xiaorong,et al. Molecular cloning and expression analysis of RbcL cDNA from the bloom-forming green alga Chaetomorpha valida (Cladophorales, Chlorophyta)[J]. JOURNAL OF APPLIED PHYCOLOGY,2014,26(4):1853-1861.
APA Deng, Yunyan,Zhan, Zifeng,Tang, Xiaorong,Ding, Lanping,Duan, Delin,&Duan, DL .(2014).Molecular cloning and expression analysis of RbcL cDNA from the bloom-forming green alga Chaetomorpha valida (Cladophorales, Chlorophyta).JOURNAL OF APPLIED PHYCOLOGY,26(4),1853-1861.
MLA Deng, Yunyan,et al."Molecular cloning and expression analysis of RbcL cDNA from the bloom-forming green alga Chaetomorpha valida (Cladophorales, Chlorophyta)".JOURNAL OF APPLIED PHYCOLOGY 26.4(2014):1853-1861.
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