Construction Of RNAi Vector Of Osvdac Genes And Expression Analysis Of The Genes In Transgenic Rice Plants

Voltage-dependent anion channel(VDAC) exist widely in fungi, plants andanimals. It play an important role in both the material transport and regulation ofenergy metablism between mitochondria and cytoplasm, and in the process ofmitochondria-mediated apoptosis.In rice genome, there were at least8vdac genes identified. In this study, nineRNA interference (RNAi) expression vectors of five osvdac genes, osvdac4, osvdac5,osvdac6, osvdac7and osvdac8were constructed by enzymatic digestion and ligationtechnology, among which, osvdac4and osvdac7RNAi vectors were transformed intoNipponbare successfully and their positive transgenic plants were obtained.Furthemore, the expression levels of eight osvdac genes in plants with down-regulatedosvdac7gene expression were comparatively analyzed by method of real-timequantitative PCR (RT-PCR). The main results are as follow:1. Nine RNAi expression vectors of five osvdac genes which named osvdac4-5,osvdac4-3, osvdac5-5, osvdac5-3, osvdac6-3, osvdac7-5, osvdac7-3, osvdac8-5andosvdac8-3were constructed by using vector of pMCG161.2. Nine RNAi expression vectors were transformed into Nipponbareindependently, but positive transgenic plants were obtained only from genetictransformation of osvdac4-3、osvdac7-5and osvdac7-3. Real-time quantitative PCR(RT-PCR) analysis demonstrated that the expression levels of those three interferedgenes in positive transgenic plants were differently down-regulated.3. The variation on expression level of eight osvdac genes in the transgenicplants with down-regulated osvdac7gene expression were analysed using real-timequantitative PCR (RT-PCR). It was observed that the down-regulation expression ofosvdac7gene was coupled with the down-regulation expression of osvdac3gene andup-regulation expression of osvdac6gene, though there was no obvious change forother members of vdac gene family.