Expression Rate of gus Driven by Eucalyptus KORRIGAN Promoter in Tissue Specific Tobacco

Abstract

KORRIGAN gene (Kor) encodes an enzyme plasma membrane bound endo-1,4-β- D-glucanase, which is member of cellulose biosynthesis genes. This gene was first discovered in Arabidopsis and later studied in other fiber crops, yet in the eucalyptus. The aim of this work focuses on Kor promoter that controls the expression of downstream gene in specific tissue and rating of expression in Tobacco (Nicotiana tabacum). Wherewith using sequence of Eucalyptus grandis genome database and extracted upstream sequence of its gene. The sequence was analyzed by in silico technique to identify for promoter characteristic and subsequently, the Kor promoter was analyzed by promoter deletion analysis. Kor promoters were designed Kor promoters, 1,587 bp. from E. grandis. It was fused to gus reporter genes and transferred to tobacco via Agrobacterium–mediation. Histochemical GUS staining of the transgenic tobacco showed that 1,587 bp. promoter conferred GUS expression in specific vascular tissues, which are in stem, petiole, leaf vain and root. The results of real time PCR analysis of 2 lines transgenic tobacco were compared with the CaMV 35S promoter showed that rate of expression about 52, 42, 11 and 9% respectively.