Construction and Screening of Genomic DNA Library of Black Gram (Vigna mungo L.) for Protease Inhibitor Genes

摘要

Plant protease inhibitors (Pis) are small proteins which represent a class of well studied compounds of plant defence genes and are abundant in storage organs such as seeds and tubers. They have been implicated in binding to proteases of insects and interfere with the digestive process, thereby proving their insecticidal activity. Like other legumes, black gram is also known to be a source of plant protease inhibitors. A genomic DNA library of Black gram cv. Uttara was constructed in Fosmid vector with more than 25 kb size inserts. Recombinant clones containing protease inhibitor gene and promoter were identified by using mung bean protease inhibitor gene as a probe. The positive clones were further confirmed by PCR using mung bean protease inhibitor, heterologous gene specific primers and subsequently sequenced and characterized by nucleotide sequence analysis.