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利用农杆菌系LBA44 0 4(pCAMBIA330 0RG)转化优良甘蓝型油菜恢复系W72 3的下胚轴节段 .pCAM BIA330 0RG含有Rss1启动子引导的雪花莲外源凝集素基因 (gna)和CaMV 35S启动子引导的除草剂抗性基因(bar) 经过两轮除草剂 (2 .5mg/Lbialaphos)筛选 (两周 /轮 ) ,除草剂抗性再生芽被转入生根培养基中生根 对根系旺盛生长的植株中所含 gna基因进行PCR分析 PCR分析证实了这些植株确为转基因植株 利用West ern印迹法对随机选择的 5株含gna基因的转基因植株的分析发现 ,其中 4株表达了 gna基因 目前正对这些表达 gna基因的转基因植株进行后代遗传分离分析
The hypocotyl segments of W72 3, an elite Brassica napus restorer line, were transformed with Agrobacterium strain LBA44 0 4 (pCAMBIA330 0RG), and pCAM BIA330 0RG contained the promoter of the gss and the CaMV 35S promoter directed by the Rss1 promoter The bootstrapping herbicide resistance bar (bar) was screened (two weeks / round) with two rounds of herbicide (2.5 mg / Lbialaphos) and the herbicide-resistant regenerated shoots were transferred to rooting medium for rooting PCR Analysis of gna Genes Contained in Plants Confirmed that These Plants Are indeed Transgenic Plants Analysis of five randomly selected transgenic plants containing the gna gene using West ern blotting revealed that four of the strains expressed the gna gene currently The transgenic plants expressing the gna gene were analyzed for offspring genetic segregation