草丁膦抗性基因（bar）在海带配子体中的稳定表达  

Stable expression of bar gene in the gametophytes of kelp （Laminaria japonica）

作　　者：张亦陈[1,2] 高江涛[1,2] 张喆[3] 姜鹏[1] 秦松[1] 

机构地区：[1]中国科学院海洋研究所,实验海洋生物学重点实验室,青岛266071 [2]中国科学院研究生院,北京100039 [3]中国海洋大学生命学院,青岛266003

出　　处：《高技术通讯》2006年第9期970-974,共5页Chinese High Technology Letters

基　　金：中国科学院知识创新工程（KZCX3-SW-215）资助项目.

摘　　要：为把海带配子体作为转基因海洋生物反应器,研究了其对除草剂草丁膦的敏感性以及温度对草丁膦处理效果的影响.结果表明：海带配子体对草丁膦敏感;统计学分析得出了草丁膦在不同温度条件下处理海带配子体的半致死浓度（LD50）及95%可信限;温差达到10℃时,相同处理时间各组内不同处理温度样品的LD50之间呈现显著差异.构建了SV40启动子驱动草丁膦抗性基因bar的表达载体pSVB,应用基因枪法转化海带配子体细胞,经40mg/L草丁膦液体培养基筛选出抗性转化子,Southern blotting结果表明bar基因已经整合到海带配子体基因组中.结论：以草丁膦作为筛选压力可以实现外源基因在海带配子体中的稳定表达.The sensitivity of kelp （Laminaria japonica ） gametophytes to PDT （phosphiothricin） at the temperature of 10℃, 15 ℃ and 20℃ was tested. The results of LD50 and its 95 % Confidence Range showed that kelp gametophytes were sensitive to PPT, and increasing the temperature could enhance the selective effect of PPT. The expression vector pSVB was constructed, which contained PPT resistance gene bar driven by SV40 promoter, and was then transformed into kelp gametophytes by particle bombardment; after screening by PPT （40mg/L） for 7d × 3times, resistance colonies were picked out. Southern blotting results showed that bar gene had integrated into the genome of bombed kelp gametophytes. PPT could be a suitable selective pressure for foreign gene stable expression in kelp gametophytes.

关 键 词：海带 配子体 生物反应器 温度 草丁膦 BAR基因 筛选标记 

分 类 号：Q785[生物学—分子生物学]