【摘要】 本研究以拟南芥DNA为模板,将罗勒烯合成酶AtTPS03基因的一个目的片段克隆到T-载体上。对克隆片段测序后,进行Blast比对,结果目的片段的序列与GenBank上报道的序列同源性达到100%。根据RNA干扰的基本原理,将目的片段正反向连到干扰载体pFGC5941查尔酮内含子的两侧,经限制性酶切和PCR扩增检测,证明已成功构建了干扰载体P-2AT03。利用农杆菌介导的浸花法转化拟南芥。收获的种子在含草丁膦的培养基上筛选抗性苗,通过对抗性苗的PCR检测,已成功获得了12株转基因苗。这些转基因苗为进一步研究罗勒烯和罗勒烯合成酶的功能奠定了良好基础。更多还原

【Abstract】 In this study, we have cloned a goal fragment of ocimene synthase AtTPS03 gene to the T-vector with Arabidopsis thaliana DNA as template. After the sequencing and blasting in GenBank, the identification between our cloned sequence and published sequence is 100%. According to the basic principles of RNA interference, the fragment was inserted to both sides of CHS intron of pFGC5941 in forwards and reverse way. The interference vector P-2AT03 was constructed successfully by restriction endonuclease analysis and PCR test. Arabidopsis thaliana was transformed by Agrobacterium tumefaciens-mediated and the recoverd seeds were screened on MS containing glufisinate ammonium. After PCR detection, 12 transgenic seedlings were obtained, which lays a good foundation for further study on the function of ocimene and ocimene synthase.更多还原