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Isolation, characterization and expression analysis of a nutritionally enhanced α-prolamin gene and protein during developing spikes of finger millet (Eleusine coracana)

Published online by Cambridge University Press:  14 September 2017

Lalan Kumar
Affiliation:
G B Pant University of Agriculture and Technology, Pantagar, Uttarakhand, India
Dinesh Pandey
Affiliation:
G B Pant University of Agriculture and Technology, Pantagar, Uttarakhand, India
Anil Kumar*
Affiliation:
G B Pant University of Agriculture and Technology, Pantagar, Uttarakhand, India
*
*Correspondence Email: [email protected]

Abstract

In the present study a combination of BLAST mediated homology search and 3′ RACE was utilized to isolate the full-length gene of Eleusine coracana alpha prolamin (Ec-α-prolamin) from finger millet. Phylogenetic analysis of Ec-α-prolamin along with related prolamin genes of different cereals and millets shows the clustering of Ec-α-prolamin in a separate group. Secondary structure prediction reveals 59.4% alpha helix structure, a structural hallmark of Ec-α-prolamin. Besides this, the protein also possesses a balanced proportion of all essential amino acids. Expression analysis based on qPCR shows increased accumulation of α-prolamin transcripts in developing grains of finger millet until attainment of seed maturity. Western blotting, using a monospecific anti-α-prolamin antibody, further confirmed the expression of a 22 kDa band in the S3 and S4 stages of developing spikes. The heterologous expression of isolated full-length Ec-α-prolamin could be potentially harnessed for making nutritionally enhanced functional food products and value-added industrial products.

Type
Research Papers
Copyright
Copyright © Cambridge University Press 2017 

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