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Cloning and characterisation of an endosperm specific glutelin B1 promoter from Sri Lankan rice (Oryza sativa L. ssp. indica) variety Bg 250

Authors:

K. Kajendran,

University of Colombo, LK
About K.
Department of Chemistry, Faculty of Science
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N. V. Chandrasekharan,

University of Colombo, LK
About N. V.
Department of Chemistry, Faculty of Science
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C. M. Hettiarachchi,

University of Colombo, LK
About C. M.
Department of Chemistry, Faculty of Science
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A. K. Nanayakkara,

University of Texas Southwestern Medical Centre, Dallas, TX, 75390, US
About A. K.

Department of Internal Medicine

 
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D.A.S. Elvitigala,

University of Colombo, LK
About D.A.S.

Department of Basic Science and Social Sciences for Nursing, Faculty of Nursing

 

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W. S. S. Wijesundera

University of Colombo, LK
About W. S. S.
Department of Biochemistry and Molecular Biology, Faculty of Medicine
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Abstract

Glutelins are the primary source of energy storage in the endosperm of rice grains. Among the glutelin promoters, Glutelin B-1 (GluB-1) is widely studied and used in transgenic rice plants to express recombinant proteins in the endosperm. In this study, three regions: 350 bp, 1308 bp and 2300 bp of the GluB-1 promoter were PCR amplified from the genomic DNA of Bg 250 rice variety. The amplified fragments were cloned into pGEM®-T Easy vector for characterisation of GluB-1 promoter. Each region of GluB-1 promoter was separately cloned into the promoterless binary vector pCAMBIA1391Z harbouring the β-glucuronidase (GUS) reporter gene. Putative transgenic plants were generated by Agrobacterium-mediated gene transformation and confirmed by PCR using nopaline synthase terminator primers. All GluB-1 promoter constructs showed expression of the GUS gene in the endosperm of T0 transgenic plant seeds. The 1308 bp GluB-1 promoter revealed the highest expression as determined by the GUS assay. This indicates the potential of this promoter for expression of recombinant proteins in rice endosperm.

 

How to Cite: Kajendran, K., Chandrasekharan, N.V., Hettiarachchi, C.M., Nanayakkara, A.K., Elvitigala, D.A.S. and Wijesundera, W.S.S., 2020. Cloning and characterisation of an endosperm specific glutelin B1 promoter from Sri Lankan rice (Oryza sativa L. ssp. indica) variety Bg 250. Journal of the National Science Foundation of Sri Lanka, 47(4), pp.383–391. DOI: http://doi.org/10.4038/jnsfsr.v47i4.9685
Published on 25 Jan 2020.
Peer Reviewed

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