Cloning and characterization of ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit (RbcS) cDNA from green microalga Ankistrodesmus convolutus

An initial study on gene cloning and characterization of unicellular green microalga Ankistrodesmus convolutus was carried out to isolate and characterize the full-length cDNA of ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit (RbcS) as a first step towards elucidating the structure of...

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Main Authors: Thanh, Tran, Chi, Vu Thi Quynh, Abdullah, Mohd Puad, Omar, Hishamuddin, Noroozi, Mostafa, Napis, Suhaimi
Format: Article
Language:English
Published: Springer 2011
Online Access:http://psasir.upm.edu.my/id/eprint/25180/1/Cloning%20and%20characterization%20of%20ribulose.pdf
http://psasir.upm.edu.my/id/eprint/25180/
https://rd.springer.com/article/10.1007%2Fs11033-011-0679-4
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spelling my.upm.eprints.251802017-09-28T07:54:24Z http://psasir.upm.edu.my/id/eprint/25180/ Cloning and characterization of ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit (RbcS) cDNA from green microalga Ankistrodesmus convolutus Thanh, Tran Chi, Vu Thi Quynh Abdullah, Mohd Puad Omar, Hishamuddin Noroozi, Mostafa Napis, Suhaimi An initial study on gene cloning and characterization of unicellular green microalga Ankistrodesmus convolutus was carried out to isolate and characterize the full-length cDNA of ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit (RbcS) as a first step towards elucidating the structure of A. convolutus RbcS gene. The full-length of A. convolutus RbcS cDNA (AcRbcS) contained 28 bp of 5' untranslated region (UTR), 225 bp of 3' non-coding region, and an open reading frame of 165 amino acids consisting of a chloroplast transit peptide with 24 amino acids and a mature protein of 141 amino acids. The amino acid sequence has high identity to those of other green algae RbcS genes. The AcRbcS contained a few conserved domains including protein kinase C phosphorylation site, tyrosine kinase phosphorylation site and N-myristoylation sites. The AcRbcS was successfully expressed in Escherichia coli and a ~21 kDa of anticipated protein band was observed on SDS-PAGE. From the phylogenetic analysis of RbcS protein sequences, it was found that the RbcS of A. convolutus has closer genetic relationship with green microalgae species compared to those of green seaweed and green macroalgae species. Southern hybridization analysis revealed that the AcRbcS is a member of a small multigene family comprising of two to six members in A. convolutus genome. Under different illumination conditions, RT-PCR analysis showed that AcRbcS transcription was reduced in the dark, and drastically recovered in the light condition. Results presented in this paper established a good foundation for further study on the photosynthetic process of A. convolutus and other green algae species where little information is known on Rubisco small subunit. Springer 2011 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/25180/1/Cloning%20and%20characterization%20of%20ribulose.pdf Thanh, Tran and Chi, Vu Thi Quynh and Abdullah, Mohd Puad and Omar, Hishamuddin and Noroozi, Mostafa and Napis, Suhaimi (2011) Cloning and characterization of ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit (RbcS) cDNA from green microalga Ankistrodesmus convolutus. Molecular Biology Reports, 38 (8). pp. 5297-5305. ISSN 0301-4851; ESSN: 1573-4978 https://rd.springer.com/article/10.1007%2Fs11033-011-0679-4 10.1007/s11033-011-0679-4
institution Universiti Putra Malaysia
building UPM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Putra Malaysia
content_source UPM Institutional Repository
url_provider http://psasir.upm.edu.my/
language English
description An initial study on gene cloning and characterization of unicellular green microalga Ankistrodesmus convolutus was carried out to isolate and characterize the full-length cDNA of ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit (RbcS) as a first step towards elucidating the structure of A. convolutus RbcS gene. The full-length of A. convolutus RbcS cDNA (AcRbcS) contained 28 bp of 5' untranslated region (UTR), 225 bp of 3' non-coding region, and an open reading frame of 165 amino acids consisting of a chloroplast transit peptide with 24 amino acids and a mature protein of 141 amino acids. The amino acid sequence has high identity to those of other green algae RbcS genes. The AcRbcS contained a few conserved domains including protein kinase C phosphorylation site, tyrosine kinase phosphorylation site and N-myristoylation sites. The AcRbcS was successfully expressed in Escherichia coli and a ~21 kDa of anticipated protein band was observed on SDS-PAGE. From the phylogenetic analysis of RbcS protein sequences, it was found that the RbcS of A. convolutus has closer genetic relationship with green microalgae species compared to those of green seaweed and green macroalgae species. Southern hybridization analysis revealed that the AcRbcS is a member of a small multigene family comprising of two to six members in A. convolutus genome. Under different illumination conditions, RT-PCR analysis showed that AcRbcS transcription was reduced in the dark, and drastically recovered in the light condition. Results presented in this paper established a good foundation for further study on the photosynthetic process of A. convolutus and other green algae species where little information is known on Rubisco small subunit.
format Article
author Thanh, Tran
Chi, Vu Thi Quynh
Abdullah, Mohd Puad
Omar, Hishamuddin
Noroozi, Mostafa
Napis, Suhaimi
spellingShingle Thanh, Tran
Chi, Vu Thi Quynh
Abdullah, Mohd Puad
Omar, Hishamuddin
Noroozi, Mostafa
Napis, Suhaimi
Cloning and characterization of ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit (RbcS) cDNA from green microalga Ankistrodesmus convolutus
author_facet Thanh, Tran
Chi, Vu Thi Quynh
Abdullah, Mohd Puad
Omar, Hishamuddin
Noroozi, Mostafa
Napis, Suhaimi
author_sort Thanh, Tran
title Cloning and characterization of ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit (RbcS) cDNA from green microalga Ankistrodesmus convolutus
title_short Cloning and characterization of ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit (RbcS) cDNA from green microalga Ankistrodesmus convolutus
title_full Cloning and characterization of ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit (RbcS) cDNA from green microalga Ankistrodesmus convolutus
title_fullStr Cloning and characterization of ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit (RbcS) cDNA from green microalga Ankistrodesmus convolutus
title_full_unstemmed Cloning and characterization of ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit (RbcS) cDNA from green microalga Ankistrodesmus convolutus
title_sort cloning and characterization of ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit (rbcs) cdna from green microalga ankistrodesmus convolutus
publisher Springer
publishDate 2011
url http://psasir.upm.edu.my/id/eprint/25180/1/Cloning%20and%20characterization%20of%20ribulose.pdf
http://psasir.upm.edu.my/id/eprint/25180/
https://rd.springer.com/article/10.1007%2Fs11033-011-0679-4
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score 13.209306