Construction and screening of subgenomic library of chickpea (Cicer arietinum L.) for legumin genes and their analysis.

ABSTRACT

Legumin storage protein genes of chickpea were identified by hybridizing restricted chickpea genomic DNA transferred on Southern blot with heterologous pea legumin cDNA probes. Southern hybridization of restricted chickpea genomic DNA with pea legumin pDUB6 and pDUB8 cDNA probes indicated presence of pea legumin homologous sequences in chickpea. About 4-5 kb EcoRI fragment was hybridized with both the probes used. DNA corresponding to this size was isolated from the gel and used for construction of subgenomic library in lambda ZAPII vector. After amplification, library was screened for presence of legumin genes in chickpea. Out of clones screened (0.1 x 10(6)), three positive clones were obtained. Restriction analysis and hybridization studies showed presence of complete structural gene in each of the identified positive clones.