Chloroplast transformation of Platymonas (Tetraselmis) subcordiformis with the bar gene as selectable marker.
Chloroplast transformation of Platymonas (Tetraselmis) subcordiformis with the bar gene as selectable marker.
复制标题
以 bar 基因作为选择标记对亚心形扁藻 (Tetraselmis) 进行叶绿体转化
DOI：
10.1371/journal.pone.0098607
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Jiang P
中科院分区：
文献类型：
--
作者：
Cui Y;Qin S;Jiang P
建立亚心形扁藻叶绿体转化技术。以gfp基因为报告基因，bar基因为选择标记，以内源片段rrn 16 S-trnI（左）和trnA-rrn 23 S（右）为叶绿体基因组重组位点，构建了亚心形紫菜叶绿体转化载体。通过粒子轰击将质粒转移到亚心形毕赤酵母中。共聚焦激光扫描显微镜显示绿色荧光蛋白定位于亚心形衣藻的叶绿体中，证实了莱茵衣藻启动子的活性。使用除草剂Basta选择用bar基因转化的细胞。通过PCR和Southern杂交分析抗性克隆，结果表明bar基因通过同源重组成功整合到叶绿体基因组中。该技术将改善这种植物的基因工程。
The objective of this research was to establish a chloroplast transformation technique for Platymonas (Tetraselmis) subcordiformis. Employing the gfp gene as a reporter and the bar gene as a selectable marker, transformation vectors of P. subcordiformis chloroplast were constructed with endogenous fragments rrn16S–trnI (left) and trnA–rrn23S (right) as a recombination site of the chloroplast genome. The plasmids were transferred into P. subcordiformis via particle bombardment. Confocal laser scanning microscopy indicated that the green fluorescence protein was localized in the chloroplast of P. subcordiformis, confirming the activity of the Chlamydomonas reinhardtii promoter. Cells transformed with the bar gene were selected using the herbicide Basta. Resistant colonies were analyzed by PCR and Southern blotting, and the results indicated that the bar gene was successfully integrated into the chloroplast genome via homologous recombination. The technique will improve genetic engineering of this alga.
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