高级检索

    毛白杨PtPCP-like基因的克隆及其遗传转化初报

    Cloning and genetic transformation of PtPCPlike from Populus tomentosa

    • 摘要: 利用滤纸吸附噬菌体PCR法,从毛白杨花芽cDNA文库中分离克隆了PtPCP-like基因cDNA全长序列,测序表明克隆得到的该序列全长595 bp,包含一个完整的开放阅读框,编码91个氨基酸;经BLAST分析发现,该基因包含与拟南芥花粉外被蛋白基因相似的序列,命名为PtPCP-like。采用RTPCR技术检测PtPCP-like基因在各个组织部位的表达模式,结果显示在毛白杨的根和雄花芽中表达丰度最高,而在雌花芽部位表达丰度最低。并且构建35S∶∶PtPCP-like植物表达载体,采用农杆菌介导法将PtPCP-like基因导入烟草中,获得了一批阳性转化植株。采用qRT-PCR技术检测PtPCP-like基因在各个转基因烟草中的表达模式,结果显示在转基因烟草中各个株系均比野生型表达量高,且不同株系间相对表达量差异显著。

       

      Abstract: To understand the genetic and molecular mechanisms of floral development in Populus tomentosa,a fulllength cDNA sequence of PtPCPlike gene was isolated from a male floral bud cDNA library of P.tomentosa,with the method of bacteriophage absorption by filter paperPCR.The sequencing results indicated that the 595 bp sequence contained a complete open reading frame, which encoded 91 amino acid residuals. BLAST analysis revealed that the gene had a similar sequence domain with Arabidopsis pollen coat protein genes, therefore it was named as PtPCPlike.RTPCR showed that PtPCPlike had higher expression abundance in male poplar floral buds and roots, and a lower abundance in female floral buds. A plant expression vector carrying 35S∶∶PtPCPlike was constructed and transformed into tobacco and poplar via Agrobacterium mediated method, a batch of positive transformation plants were obtained. Expression pattern of PtPCPlike in different tobacco lines was examined using qRTPCR. Results showed that PtPCPlike was expressed higher in all transgenic lines than in wide type, and significant expression differences were seen among different transgenic lines.

       

    /

    返回文章
    返回