Tools for chloroplast transformation in Chlamydomonas:: expression vectors and a new dominant selectable marker
Tools for chloroplast transformation in Chlamydomonas:: expression vectors and a new dominant selectable marker
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DOI：
10.1007/s004380051184
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发表时间：
2000-04-01
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影响因子：
--
通讯作者：
Purton, S
中科院分区：
文献类型：
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作者：
Bateman, JM;Purton, S
莱茵衣藻叶绿体基因的反向遗传研究由于缺乏合适的叶绿体转化选择标记而受到阻碍。我们构建了一系列外源基因在衣藻叶绿体基因组中靶向插入和表达的载体。利用这些载体，我们基于细菌基因aphA-6开发了一种新的选择性标记，该标记编码氨基糖苷磷酸转移酶。apha -6,4-6标记允许在含有卡那霉素或阿米卡星的培养基上直接选择转化子。该标记可用于灭活或修饰特定的叶绿体基因，并可作为基因表达的报告者。该标记的存在使衣藻叶绿体基因组的序列转化成为可能。
Reverse-genetic studies of chloroplast genes in the green alga Chlamydomonas reinhardtii have been hampered by the paucity of suitable selectable markers for chloroplast transformation. We have constructed a series of vectors for the targeted insertion and expression of foreign genes in the Chlamydomonas chloroplast genome. Using these vectors we have developed a novel selectable marker based on the bacterial gene aphA-6, which encodes an aminoglycoside phosphotransferase. The aphA-6,4-6 marker allows direct selection for transformants on medium containing either kanamycin or amikacin. The marker can be used to inactivate or modify specific chloroplast genes, and can be used as a reporter of gene expression. The availability of this marker now makes possible the serial transformation of the chloroplast genome of Chlamydomonas.