破囊壶菌Δ~4-脂肪酸脱饱和酶基因5'端侧翼区域的克隆与鉴定Cloning and Functional Analysis of 5' Flanking Region of Δ~4-Desaturase Gene from Thraustochytrium
江贤章;陈金卿;田宝玉;高媛媛;舒正玉;李欣;蓝灿华;黄建忠;
摘要(Abstract):
应用衔接头PCR(LA-PCR)技术,扩增得到约1000 bp的海洋破囊壶菌Δ4-脱饱和酶基因5’端侧翼区域的单一产物.对该产物测序,经启动子软件与序列比对分析,发现该序列(Genkbank登录号EU074209)具有真核启动子序列的基本结构特征,含有TATA盒、CAAT盒、INR等元件.将扩增得到的脱饱和酶基因5’端侧翼序列亚克隆到含有绿色荧光蛋白(GFP)报告基因的质粒pGlow-TOPO中,构建重组表达质粒,转化大肠杆菌细胞.荧光显微观察大肠杆菌阳性转化子发出荧光,侧翼序列含有启动子功能得到确认.
关键词(KeyWords): 破囊壶菌;脱饱和酶;衔接头PCR;启动子;绿色荧光蛋白
基金项目(Foundation): 国家自然科学基金资助项目(30370028);; 福建省自然科学基金重大项目(2003F005);; 福建省教育厅基金资助项目(JB07079);; 福建省发改委重大项目([2004]477);; 福建省科技厅重大项目(2005Q007);; 福建省青年人才基金资助项目(2008F3036)
作者(Authors): 江贤章;陈金卿;田宝玉;高媛媛;舒正玉;李欣;蓝灿华;黄建忠;
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