甘蓝型油菜二酰甘油酰基转移酶基因微藻真核表达载体的构建

Construction of eukaryotic expression vector for microalga using cloned DGAT genes from Brassica napus

目的:构建二酰甘油酰基转移酶(DGAT)的微藻真核表达载体。方法:采用RT-PCR从甘蓝型油菜中扩增得到DGAT1和DGAT22个基因cDNA编码区序列,分别连接到载体pMD18-TSimple上,经测序及与已知的DGAT1和DGAT2序列比对,将鉴定正确的基因用于微藻真核表达载体pSV40DGAT1/CaMVBar和pSV40DGAT2/CaMVBar的构建。结果:克隆到的甘蓝型油菜DGAT1和DGAT22个基因长度分别为1512和1026bp,同源性分别为99%和98%,构建了微藻真核表达载体pSV40DGAT1/CaMVBar和pSV40DGAT2/CaMVBar。结论:2个微藻真核表达载体构建成功。

Aim：To construct the diacylglycerol acyltransferase（DGAT） eukaryotic expression vector for microalga.Methods：DGAT1 and DGAT2 were obtained from Brassica napus through RT-PCR and then cloned into vector pMD18-T Simple.The cDNA fragments indentified correctly by sequencing were subcloned into an eukaryotic expression vector to construct pSV40DGAT1/CaMVBar and pSV40DGAT2/CaMVBar.Results：The cloned cDNA sequences were 1 512 and 1 026 bp with hemology of 99% and 98%,respectively,and were utilized for constructing eukaryotic expression vectors pSV40DGAT1/CaMVBar and pSV40DGAT2/CaMVBar.Conclusion：The two eukaryotic expression vectors for microalga have been successfully constructed.