Identification Of Over-expression OsVDAC3 Transgenic Plants And Construction Of OsVDAC1 Prokaryotic Expression Vector

VDAC(voltage-dependent anion channel)protein is a kind of pore protein which widely exists in various eukaryotic species.It regulates the import and export of material through the change of voltage.Eight different vdac genes have been identified in the rice genome in our laboratory.Compared to the wild type YTB,the transgenic plants with overexpressed OsVDAC3 were more sensitivity to the cold and salt tolerance.Also we got a batch of seeds of YTB plants with overexpressed QsVDAC3.In this experiment,we expect to obtain some positive plants which are suitable for further study from this batch of materials,and construct the prokaryotic expression vector for OspVDACI.The main results are as follows:1.The seeds of the YTB with OsVDAC3 overexpressed were cultured and then the genomic DNA was isolated.Firstly the Hyg positive plants were identifed by PCR.Within the Hyg positive plants the Rf5 and Rf6 negative plants were identified.And at same time if it is positive for OrfH79 and Orf216 the receptor for transgenic is YTA,if not the receptor is YTB.After screening and review,there were dozens of OsVDAC3 overexpressed YTB plants identified.but no YTA plants.2.The ORF of OsVDAC1 was 825 bp and code 274 amino acid residues through bioinformatics anslysis.The ORF was amplified from Nipponbare cDNA,and the recombinant plasmid pET-32a-O5sVDAC1 was constructed.The results of SDS-PAGE and Western blot showed that OsVDAC1 was induced successfully by IPTG.