Construction and identification for differential expression cDNA subtractive library in HT-29 cell lines induced by DADS

Objective: To construct differential expression cDNA subtractive library of human colon carcinoma HT-29 cell lines induced by DADS(diallyl disulfide,DADS),to find and cloning DADS specific target gene.Methods: Human colon carcinoma HT-29 cell was processed with 120μmol/L DADS,Poly A+ RNA were isolated from colon carcinoma HT-29 cells treated for trial group and control group respectively.Differential expression cDNA subtractive library was constructed by using the suppression subtractive hybridization(SSH),then used it to transfect E.coli strain JM109,the library was amplified with the transfected E.coli strain JM109.Results: Library of human colon carcinoma HT-29 cells induced by DADS subtractive library with high subtractive efficiency was set up successfully.The amplified library contained 500 clones.100 clones were selected randomly to make plasmid,with PCR ampification showed that all plasmids in the clones contained 150～1300 bp insert sequences;which might contain high specific sequences of target.Conclusion: The cDNA subtractive library of human colon carcinoma HT-29 cells is constructed,which lays a foundation for searching and cloning specific target gene of DADS.