Construction of cDNA library from peanut pod by using suppression subtractive hybridization

To isolate peanut pod specially expressed genes,a cDNA library was constructed by using suppression subtractive hybridization(SSH).For subtractive hybridization,cDNA from the kernel of peanut was used as Driver,and cDNA from the pod of peanut was used as Tester.After two times of subtractive hybridization and two times of nested PCR,the products of last PCR amplification were inserted into pGEM-T Easy vectors and be transformed into the E.coli,a cDNA library of peanut pod was successfully constructed.89.2% of the colonies were white.The positive recombinants were confirmed by PCR method,83.5% of them showed single band.Those cDNA in the library can be used to isolate differentially expressed genes and promoters of peanut pod,also can be further studied for the purpose of understanding the molecular mechanism in the specially expression of pod genes.