【作者】 谷立坤；

【导师】 贾新成； 陈红歌；

【作者基本信息】 河南农业大学 ， 微生物学， 2004， 硕士

【摘要】 本文以TPS1基因的CDNA,质粒pWM101为研究对象，运用基因克隆技术，成功构建了植物表达载体pWM102。将质粒PWM102转化农杆菌菌株EHA105，经kana霉素，Rif霉素筛选，PCR鉴定，获得含有重组质粒PWM102的农杆菌。通过对诱导培养基，分化培养基中2，4-D浓度调整，得到了一组对成熟胚诱导与分化较好的培养基，并确定以小麦温6的成熟胚与幼胚作为基因转化的外植体，利用农杆菌介导的方法，将海藻糖-6-磷酸合成酶基因（TPS1）导入小麦温6，幼胚与成熟胚产生的愈伤组织中，在含有20-60mg/l潮霉素的继代培养基上，分别获得了一批抗性愈伤组织，经过组织分化，获得了一批再生苗。并且在其中四株再生苗中检测到TPS1基因，结果表明四株扩增出TPS1基因片段的植株为转基因植株。更多还原

【Abstract】 The fragment of tpsl was digested by two restriction enzyme Kpnl/Xbal ,and then was successfully joined with Pwml01 plasmid which was digested by Kpnl/Xbal too.The outcome was expression vector Pwml02,which wastransformed into Agrobacterium tumefaciens EHA105.By changing the concentration of the 2,4-D,we get a good inducing and differentiation medias, and deciding using the mature and immature embryos of Wen 6 as explants.Agrobacteriurn mediates transforming the TPS1 gene into the callus forming by mature and immature embryos of wen 6.Then we get some resistencing callus which could growing on medias containing 20-60mg/1 hygromycin.We get some regernerating wheats and four of them are positive by PCR analysis.These four plants were indentified to bethe transgenic wheats.更多还原