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IJBT Vol.05 [2006] >
IJBT Vol.05(1) [January 2006] >
| 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 Code: | Int. 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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