[反]-β-法尼烯(EβF)合成酶基因在玉米中的转化与筛选

Transformation of( E)-β-farnesene Synthase Gene into Maize and Preliminary Screening

为获得转基因抗虫玉米,将[反]-β-法尼烯(EβF)合成酶基因导入玉米,并进行鉴定和筛选。在[反]-β-法尼烯(EβF)合成酶基因的两端分别添加Nco I和Bam H I酶切位点,并进行人工合成。将EβF合成酶基因与植物表达载体p FGC5941连接,经双酶切和测序鉴定。结果表明:重组表达载体p FGC5941-EβF构建成功;载体p FGC5941-EβF转化农杆菌EHA105后用玉米芽尖侵染法将EβF合成酶基因导入玉米自交系郑58中,对转化植株进行除草剂抗性筛选以及EβF合成酶基因和bar基因的PCR鉴定后,共得到18株转基因阳性植株。

To obtain insect-resistant transgenic maize, （E）-β-farnesene synthase gene was transferred into maize, which was identified and screened. The Nco I and Bam H I enzyme cutting sites were separately added at both ends of the （E） -β-farnesene synthase gene. This sequence was artificially synthesized. The desired gene was ligated into vector pFGC5941 by double enzyme digestion and DNA ligase to con- struct ecombinant expression vector pFGC5941-EβF, which were sequenced and identified by double enzyme digestion. The result showed that the vector pFGC5941-EβF was successfully constructed, Then pFGC5941-EβF was transformed into Agrobacterium tumefaciens EHA105 that infected shoot apical growing point of maize Zheng58. The transgenic maize plants were identified by herbicide-resistant and PCR amplification for bar and EβF synthase genes, and a total of 18 positive transgenic maize plants were obtained.