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

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Abstract

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.

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Acknowledgments

This work was supported by the MOSTI grant number 02-01-04-SF0041 from the Ministry of Science, Technology and Innovation, Malaysia. The first author is indebted to the Rubber Research Institute of Vietnam on the financial support for the postgraduate scholarship.

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Authors and Affiliations

  1. Department of Cell and Molecular Biology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400, UPM Serdang, Selangor Darul Ehsan, Malaysia

    Tran Thanh, Vu Thi Quynh Chi, Mohd Puad Abdullah & Suhaimi Napis

  2. Department of Biology, Faculty of Science, Universiti Putra Malaysia, 43400, UPM Serdang, Selangor Darul Ehsan, Malaysia

    Hishamuddin Omar & Mostafa Noroozi

  3. Rubber Research Institute of Vietnam, 236Bis Nam Ky Khoi Nghia Street, District 3, Ho Chi Minh City, Vietnam

    Tran Thanh & Vu Thi Quynh Chi

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  1. Tran Thanh
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  2. Vu Thi Quynh Chi
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  3. Mohd Puad Abdullah
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  4. Hishamuddin Omar
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  6. Suhaimi Napis
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Correspondence to Suhaimi Napis.

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Thanh, T., Chi, V.T.Q., Abdullah, M.P. et al. Cloning and characterization of ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit (RbcS) cDNA from green microalga Ankistrodesmus convolutus . Mol Biol Rep 38, 5297–5305 (2011). https://doi.org/10.1007/s11033-011-0679-4

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  • DOI: https://doi.org/10.1007/s11033-011-0679-4

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