坛紫菜转鲎素基因的初步研究  

Preliminary Study on Transfer Tachyplesin I Gene into Porphyra haitanensis

作　　者：吴顺章[1] 李博文[1] 赵扬[1] 陈奕欣[1] 

机构地区：[1]厦门大学生命科学学院

出　　处：《厦门大学学报（自然科学版）》2008年第4期567-570,共4页Journal of Xiamen University：Natural Science

基　　金：福建省自然科学基金(B0510004);福建省重大农业科技项目(2001Z017)资助

摘　　要：以坛紫菜为材料,利用电穿孔法将构建的含有鲎素基因tachyplesinI的同源重组表达载体pSV40-HR-MAR-tachyplesinI导入坛紫菜叶状体体细胞.并通过PCR、Southern杂交等分子生物学检测方法对导入后的细胞进行检测,结果表明构建的以坛紫菜18S rDNA的460、1200bp的序列作为外源基因3′端和5′端同源重组序列,同时以家蚕的MARs序列作为增强表达序列的同源重组表达载体导入坛紫菜叶状体体细胞后,鲎素基因tachyplesinI能整合到坛紫菜叶状体体细胞内.通过Northern点杂交检测的结果表明,鲎素基因能在坛紫菜体细胞内瞬时表达.本研究构建的鲎素基因同源重组表达载体可用于相关大型海藻的转基因研究中.In this study,recombination vector pSV40-HR-MAR-tachyplesin Including Matrix attachment regions and 18S rDNA for targeted homologous recombination was constructed and introduced into Porphyra haitanensis cells with the method of electroporation. Then integration of tachyplesin I in the P. haitanensis cells was confirmed with both PCR and southern blotting. The results showed that tachyplesin I was introduced successfully into P. haitanensis cells and integrated. Furthermore, northern dot blotting was performed and the result showed that tachyplesin I expressed at the level of RNA in the transgenic P. haitanensis cells. These results indicated that the strategy was useful for transformation,integration and expression of tachyplesin I within P. haitanensis cells. The recombination vector for expression tachyplesin I could be applied in the interrelated research of transgenic large seaweed.

关 键 词：坛紫菜 鲎素 转基因 

分 类 号：Q953[生物学—动物学]