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Title: Isolation and characterization of gene encoding vicilin (7S) protein from cDNA clones of immature seeds of pigeon pea [Cajanus cajan (L.) Millsp.]
Authors: Gupta, Subodh
Verma, A K
Chand, Laxmi
Keywords: cDNA
gene
pigeonpea
storage protein
vicilin (7S)
Issue Date: Jan-2006
Publisher: CSIR
IPC CodeInt. Cl.7 C12N15/11
Abstract: cDNA library was constructed in lgt 11 expression vector from mRNA of immature seeds of pigeonpea.  This cDNA was screened with specific non-radioactive, DIG-labelled heterologous pea vicilin cDNA probe (pRC-758). The positive vicilin (7S) encoding cDNA clones were isolated. One of the vicilin cDNA clones showed insert size of ~1.3 Kb, when analyzed by PCR using lgt 11 forward and reverse primers. This PCR product was subcloned in pUC-18 vector for confirmation of gene and product by Southern and Western hybridizations, respectively. The clone was named as pSL-1 and sequenced with M13 universal forward and reverse sequencing primers. The partial nucleotide sequence (1341 bp) has been indexed in NCBI gene bank with accession number AF348366. The complete gene has 1417 base pairs with 972 bp coding sequence. Hence, the predicted polypeptide chain of this gene was determined, which contained 323 amino acids. After post translation modification, the predicted polypeptide has 310 amino acids long with approximately mol wt of 34.1 kDa. This gene encoding vicilin (7S) protein provides the basic understanding of the gene structure of pigeonpea storage proteins.
Page(s): 32-36
ISSN: 0975-0967 (Online); 0972-5849 (Print)
Source:IJBT Vol.05(1) [January 2006]

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