Antiserums Preparation Of VpSBP3and VpSBP16, Identification And Analysis Of ZHD Gene Family, From Grape

SQUAMOSA PROMOTER BINDING PROTEIN (SBP) is a class of transcription factor,which is specific to plants. It has been shown that, SBP transcription factor family has animportant role in many aspects of biological functions. On this study, we try to test and verifythe expression in E. coli and function of SBP genes by experimental techniques includingprokaryotic expression, polyclonal antibodies preparation and Western blot, et al, so that thiscould supply the further function research of SBP gene family. Some primary results of thisstudy are as follows:(1) After translating the recombinant prokaryotic expression vectorspGEX-6p-1/VpSBP3and pGEX-6p-1/VpSBP16into cells of E. coli Rosetta, respectively, thefusion proteins were expressed by induction of IPTG.(2) The GST-VpSBP3fusion proteinexpression was enhanced, resulted the optimizing of induction IPTG concentration.(3) TheGST-VpSBP3and GST-VpSBP16fusion proteins were purified by the electrodialysis,respectively, then the purified products were identified by the SDS-PAGE electrophoresis.(4)The polyclonal antiserums of VpSBP3and VpSBP16were prepared by immunized the rabbitwith their purified fusion proteins, respectively. Then, the polyclonal antiserums were testedby Western blot, showing a good specific antigen-antibody recognition feature.(5) TheWestern blot analysis of VpSBP3and VpSBP16protein in various organs and in fruit atdifferent developmental stages from V. quinquangularis ‘shang-24’ suggest that they mayhave an important function in the vegetative and reproductive phase transitions and the fruitmature stages.Plant zinc finger-homeodomain (ZHD) genes encode a family of plant specialtranscription factors. On this study, all ZHD genes were identified in grape genome byBioinformatics approaches. Phylogenetic and synteny analysis were used to investigate theevolution of the ZHD gene family. We further analyzed the expression profiles of grape ZHDgenes under various abiotic, biotic stresses and xenobiotics treatments throughsemi-quantitative RT-PCR and real-time PCR. Some primary results of this study are asfollows:(1) We identified a total of13ZHD genes (VvZHD) in the grape genome that werefurther classified into at least six groups, and were named based on their genome location.(2)Genome synteny analysis revealed that a number of VvZHD genes were present in the corresponding syntenic blocks of Arabidopsis, indicating that they arose before thedivergence of these two species.(3) Gene expression analysis showed that the identifiedVvZHD genes displayed distinct spatiotemporal expression patterns, and were differentiallyregulated under various stress conditions and xenobiotics treatments, suggesting that the grapeVvZHDs might be also involved in plant response to a variety of biotic and abiotic insults.