毛白杨PtPCP-like基因的克隆及其遗传转化初报
Cloning and genetic transformation of PtPCPlike 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RTPCR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。
-
关键词:
- 毛白杨 /
- 烟草 /
- qRT-PCR /
- PtPCP-like
Abstract: To understand the genetic and molecular mechanisms of floral development in Populus tomentosa，a fulllength cDNA sequence of PtPCPlike gene was isolated from a male floral bud cDNA library of P.tomentosa，with the method of bacteriophage absorption by filter paperPCR.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 PtPCPlike.RTPCR showed that PtPCPlike 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∶∶PtPCPlike was constructed and transformed into tobacco and poplar via Agrobacterium mediated method, a batch of positive transformation plants were obtained. Expression pattern of PtPCPlike in different tobacco lines was examined using qRTPCR. Results showed that PtPCPlike was expressed higher in all transgenic lines than in wide type, and significant expression differences were seen among different transgenic lines.-
Keywords:
- Populus tomentosa /
- tobacco /
- qRT-PCR /
- PtPCP-like
-
计量
- 文章访问数: 2407
- HTML全文浏览量: 119
- PDF下载量: 43
下载:
