Molecular cloning of rolC gene homologue from lettuce(Lactuca.) and identification of rolC gene expression in plant tissues

Abstract

rolC gene homologue from Lettuce was cloned and its expression was confirmed in plant tissues. Firstly, the homologous region in several plant chromosomal DNA was detected by DNA slot blot hybridization. Lettuce total RNA was analyzed by norther n blot analysis and revealed that the rolC gene was being expressed. Poly(A)+ RNA was selected from total RNA, analyzed by RNA slot blot analysis, and revealed that the rolC gene does not be expressed after about 6 week. Then, Lettuce genomic DNA was digested with HindIII and analyzed by southern blot hybridization. DNA fragments of 4-6 kb range were subcloned to pUC19 and screened by colony hybridization. 4 colones were mapped with several restriction enzyme. After localization of pKC01, Avaii fragment was subcloned to pSELECT-1.