Isolation of fruitwall-specific gene from cotton plant (Gossypium hirsutum)

Viral clones containing DNA specific for bollwall tissues of glandless cotton plants (Gossypium hirsutum) were isolated. Subtractive Southern hybridization procedures of cotton nuclear genomic library was the main screening strategy using bollwall and leaf cDNA probes. Viral DNA of the specific clones were analyzed and a fruitwall-specific gene from cotton plants was identified. Southern hybridization analysis for the viral genomic clones emphasized the predicted results in obtaining fruitwall-specific gene on a genomic fragment insert, and identified the presence of the bollwall-specific gene in the cotton nuclear genome. Northern hybridization analysis verification for tissue specificity indicated the presence of 1.2 kbp fruitwall-specific gene only in bollwall tissue and tomato fruitwall tissue. The fruitwall-specific gene was located on 2.7 kb fragment which was subcloned and proved its specificity in subtractive Southern hybridization experiment. Genomic Southern hybridization experiment showed the localization of the gene of concern on an intact 2.7 kb genomic fragment.