绞股蓝RIP基因双子叶植物表达载体的构建及其对烟草叶盘的转化

Construction of Dicotyledon Expression Vector Containing cDNAEncoding Ribosome-inactivating Protein from Gynostemma pentaphyllum and Transformation into Tobacco

将绞股蓝(Gynostemmapentaphyllum)核糖体失活蛋白(Ribosome-InactivatingProtein,RIP)cDNA序列(Gen Bank登录号AY279219)插入到pKYLX71∶35S2植物表达载体的HindIII/SacI位点处,构建了适于在双子叶植物中表达的载体,并经三亲交配法转入土壤农杆菌LBA4404,用于转化烟草品种K-326,分子鉴定表明绞股蓝RIP基因已经整合到再生烟苗的基因组中并发生了转录。

The cDNA sequence (825 bp in length, GenBank accession number AY279219) encoding ribosome-inactivating protein from Gynostemma pentaphyllum was inserted into the HindⅢ/SacⅠ site of pLYLX71∶35S^2. an expression vector for dicotyledon. The Agrobacterium tumefaciens LBA404 strain harboring the cDNA via trimated transformation was used to infect the tobacco variety K326. The results of molecular detection demonstrated that the target cDNA had been integrated into the genome of regenerated tobacco shoot and transcripted.