Cloning of Chalcone Reductase GmCHR Gene and Its Expression in Tobacco

GmCHR gene sequence was isolated from soybean varietyjinong 28by RT-PCR technique.The 918 bp open reading frame was selected in that sequence.On the basis of pBI121 carrier,a GmCHR recombinant botany expression vector driven by 35 S promoter was constructed and transferred into tobacco.Seventeen plants of kan resistant transgenic tobacco were identified by molecular biology method of PCR and Southern blotting.Tested positive transgenic tobacco was performed with GUS activity assay.Expression value of the gene in different plants was determined by real-time fluorescent quantitative PCR and the content of isoliquiritigenin catalyticed product by the enzyme was identified by HPLC technology.The results indicated that GmCHR gene had been integrated into the tobacco genome and expressed successfully.The expression value of the gene in the leaves of different transgenic plants was significantly different.Average content of isoliquiritigenin catalyzed by enzyme in tobacco leaf tissue was(7.528±0.018) μmol /g,but isoliquiritigenin was not detected in empty vehicle transfected tobacco.