Genetic transformation and expression detection of tobacco transformed by BtCry1Ac and BADH bivalent gene
Tobacco was transformed by plant transformation vector p209-BtCry1Ac-BADH through Agrobacterium-mediated,the complete regenerated plants were obtained by screening of Kanamycin.4 transgenic lines that detected BtCry1Ac and BADH were obtained after PCR detection.Fluorescence quantitative PCR indicated that two target genes were expressed at the transcriptional level in 3 lines that detected all the target genes,and there were some difference among these lines.However,the expression of BADH in a line was not detected.ELISA detection of toxic protein showed that the expression of toxic protein in 3 lines screened by fluorescence quantitative PCR was detected,with the maximum content of toxic protein of 414.63 ng/g.And the expression of toxic protein in the line with detected BtCry1Ac was detected.The indoor insects-resistance test showed that only 2 lines in the 3 transgenic lines showed the resistance against Prodenia litura (Fabricius),with the maximum corrected mortality of 38.2％.The others showed no obvious resistance to the larve.Additionaly,the results of the plantlets salt-tolerance test in 2 lines selected revealed that there were no significant differences between the transgenic lines and the control.In the study,BtCry1Ac gene and BADH gene of some lines had been integrated into the plant genome by PCR,but the quantitative PCR and ELISA showed that although the gene into the plant genome no expression was detwecxted,which was possiblely due to the gene silencing.
NETL
NSTL
万方数据
维普
