疱疮丙酸杆菌亚油酸异构酶基因转化亚麻芥

Transformation of Camelina sativa with a Linoleic Acid Isomerase Gene from Propionibacterium acnes

亚油酸异构酶可特异性地催化亚油酸转化为活性共轭亚油酸异构体。本研究采用PCR技术从疱疮丙酸杆菌GIM1.243中扩增出亚油酸异构酶基因PAI,片段长1275bp。按照目标宿主亚麻芥的密码子偏好性对该基因部分序列进行优化,命名为CaPAI。借助双T-DNA表达载体pSB130,用菜豆种子特异性启动子PhaP替换原有的CaMV35S启动子,驱动CaPAI,终止子替换成菜豆的终止子PhaT;另一个T-DNA保留其原有的表达框,其中含有潮霉素筛选标记基因。通过农杆菌介导的FloraDip转化法,将构建好的高效植物表达载体pSB130-PhaP-CaPAI-PhaT转入亚麻芥中,抗性筛选和PCR鉴定显示CaPAI基因已整合进亚麻芥基因组中。

Linoleic acid isomerase can specifically catalyze the linoleic acid（LA） and then convert it to conjugated linoleic acid（CLA） isomer.In this research,the LA isomerase gene PAI with fragment length of 1 275 bp was amplified from Propionibacterium acnes GIM1.243 by PCR.The partial sequence of the gene was optimized according to the codon preference of the target host Camelina sativa,which named as CaPAI.With the help of the double T-DNA expression vectors p SB130,the original CaMV35 S promoter was replaced with the bean seedspecific promoter PhaP,the CaPAI was driven,and the terminator was replaced with the terminator PhaT of bean.and there was a selected marker gene HYG in the other T-DNA region.The constructed vector pSB130-PhaP-CaPAIPhaT was transferred into Camelina sativa by the Agrobacterium-mediated flora dip method.Antibiotics screening and PCR analysis showed that CaPAI gene had been integrated into Camelina sativa genome.