FAE1启动子的克隆和转γ-TMT基因大豆的获得  

Cloning of Fatty Acid Elongase 1 Promoter and Obtainment of γ-Tocopherol Methyltransferase Transgenic Soybean

作　　者：张明[1] 陈德富[1] 王永芹[1] 王绘砖[1] 陈喜文[1] 

机构地区：[1]南开大学生命科学学院,天津300071

出　　处：《南开大学学报（自然科学版）》2010年第6期35-41,共7页Acta Scientiarum Naturalium Universitatis Nankaiensis

基　　金：天津市自然科学重点基金(07JCZDJC03800);国家自然科学基金(31070273)

摘　　要：脂肪酸链延长酶组分1(FAE1)是广泛存在于植物中催化脂肪酸碳链延长的酮脂酰CoA合成酶,该反应是脂肪酸碳链延长的限速步骤,在油料种子胚中特异性表达.以拟南芥基因组DNA为模板,设计引物,扩增到该基因的启动子片段.序列分析表明,该片段长度为943 bp,与GenBank注册序列AF355982达99.5%同源,并含有与之完全相同的种子特异性表达元件,据此认定它具有种子特异性启动活性.将其与甘蓝型油菜γ-生育酚甲基转移酶(γ-TMT)基因连接,构建成表达载体pFAE1-TMT,通过农杆菌介导的方法转化大豆胚尖,获得28株卡那霉素抗性筛选植株,进一步PCR检测获得2株转γ-TMT基因植株.Fatty acid elongase 1 generally exists in plants＇ seed and plays an important role in the production of very-long-chain fatty acid.It acts as a rate-limiting enzyme and expresses exclusively in developing embryo.A fragment of its promoter was amplified from Arabidopsis genomic DNA.Sequence analysis indicated the fragment is 943 bp and had 99.5%homologies compared with the registered sequences in GenBank（access No.AF355982） and contained the completely identical elements for seed-specific expression as it and was supposed to have the activity to direct gene downstream express exclusively in seed.It was recombined with theγ-TMT gene from Brassica napus and a plant expression vector pFAE1-TMT was constructed and then transformed into soybean embryonic tips using Agrobacterium tumefaciens-mediated method.Twenty-eight kanamycin resistant plantlets were obtained,and two of them were confirmed with the integration ofγ-TMT gene by genomic DNA PCR.

关 键 词：脂肪酸链延长酶1(FAE1) 启动子 γ-生育酚甲基转移酶(γ-TMT) 种子特异性表达载体 转基因大豆 

分 类 号：Q943[生物学—植物学]