Interactor Screen Of OsVDAC4 And OsVDAC5 Promoter Expression Pattern Analysis

Voltage-dependent anion channels(VDACs)are ubiquitous mitochondrial pore proteins that control the transport of metabolites and inorganic salts,maintain homeostasis in the environment,and are involved in apoptosis.There were eight kinds of VDAC proteins in rice identified before.In this experiment the interaction proteins with OsVDAC4 are screened and the promoter of OsVDAC5 is preliminary studied.The main results are as follows:1.The OsVDAC4 full-length sequence was ligated into two bait protein vectors pBT3-N and pBT3-SUC.The results of yeast double-hybrid technique of membrane protein showed that pBT3-SUC-OsVDAC4 could be used for screening the interaction-protein with OsVDAC4.There were 4 proteins interacted with OsVDAC4 screened from young panicle cDNA library in rice YTB by yeast split-ubiquitin system.The point-to-point hybridization experiments demonstrated that only LOC_Os02g06640.1 and LOC_Os06g51220.2 interacted with OsVDAC4 and tentatively named OsV4IP1 and OsV4IP2.Bioinformatics analysis revealed that OsV4IP1 has 14 or 22 transmembrane or no transmembrane.OsV4IP2 protein has no transmembrane structure.Both OsV4IP1 and OsV4IP2 are located on the nucleus.2.The upstream 4,500 bp from ATG of OsVDAC5 was divided into four 5'excalation fragments pB,pC,pD and pE through bioinformatics analysis.The four fragments were ligated into the binary expression vector DX2181,respectively,and the transgenic rice plants were obtained mediated by agrobacterium.Currently only there were positive plants expressing GFP with pC,pD,and pE.The results from observations of fluorescent in the transgenic plants showed that pC,pD and pE expressed in the sclerenchyma,endodermis,stele of root.pC and pD expressed in the epidermis and stele of leaf,the phloem,ligule,leaf node of sheath,shell,stamen and pistil.There were no expression in the cortex parenchyma of root,mesophyll and sheath internal.