Cloning And Analysing For SBP Gene Sequence In Chinese Wild Vitis Species

Uncinula necator is one of fungous diseases that cause severe damage on grape.There is abundant resistance in Chinese wild Vitis species. Utilizing these excellent germplasm to breed new grape varieties,which have not only good cultivated characters but also disease resistance,is along-time target of disease resistance breeding in grape. In order to study resistance uncinula necato gene, our study grou have gained 10 resistanced ncinula necato cDNA fragments by the technology of mRNA differential display reverse transcription-PCR, DDRT-PCR and anlysised by BLAST show that the T11GG/B0324-292 cDNA fragment and Arabidopsis thaliana SBP have high aligenment.so the the T11GG/B0324-292 cDNA fragment is short partial cDNA fragment of Chinese Wild vitis baihe35-1VpSBP,we adopt rapid amplifieation of cDNA ends (RACE). and bioinformatics soft Blast searches alignment in DNA database on thebase of T11GG/B0324-292 cDNA fragment,we gained these results:1 Cloning and analysis of VpSBP cDNA and DNA sequence (VpSBP) from Baihe-35-1: Cloning and analysis 3'cDNA of VpSBP from Baihe-35-1 on the base of T11GG/B0324-292 cDNA fragment ,we gain a partial cDNA fragment,its length is 1177bp, 3'cDNA of VpSBP has highly identity to some plants'cDNA,such as SPL1,SPL2.2 Cloning and analysis of VpSBP cDNA and DNA sequence (VpSBP)from Baihe-35-1 The full length cDNA sequence of VpSBP was cloned using RACE and specific primer and was temporarily designated as VpSBP, its full length is 2566bp and have integrity open reading frame but also encode 640 amino acid ; .The deduced amino acid sequences of VpSBP shared highly identity to some plants'SBP gene between 74%～87%.3 To gain the high purity and integrity RNA.we used the modified SDS/Phenol. The modified SDS/Phenol method can extract undegraded total RNA from mature leaf and young leaf, FollowingThis efficient procedure,we routinely obtained the high purity and integrity RNA, The ratio of A260/A280 =1.8-2.0. The RNA is highly stable and sufficiently pure for DDRT-PCR,RACE etc.