CHLOROPLAST TRANSFORMATION IN CHLAMYDOMONAS WITH HIGH-VELOCITY MICROPROJECTILES
CHLOROPLAST TRANSFORMATION IN CHLAMYDOMONAS WITH HIGH-VELOCITY MICROPROJECTILES
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DOI：
10.1126/science.2897716
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发表时间：
1988-06-10
期刊：
影响因子：
56.9
通讯作者：
SANFORD, JC
中科院分区：
文献类型：
--
作者：
BOYNTON, JE;GILLHAM, NW;SANFORD, JC
用包裹有野生型基因的克隆叶绿体DNA的高速钨微弹轰击莱茵衣藻叶绿体atpB基因的三个突变体，永久恢复了该藻的光合作用能力。在其中一个突变体的大多数转化子中，2.5kb缺失的片段通过同源替换事件恢复到正常大小；在大约25%的转化子中，恢复的限制性片段比野生型的片段小或大50到100个碱基对。当第一次检查时，这个突变体的大约四分之一的转化子含有未整合的供体质粒。在连续液体培养65代后，该质粒在4个不同的转化子中持续存在，但在选择培养基板上保存的所有转化子中均丢失。恢复的野生型atpB基因作为叶绿体基因组的组成部分保留在所有转化子中，并正常表达和遗传。
Bombardment of three mutants of the chloroplst atpB gene of Chlamydomonas reinhardtii with high-velocity tungsten microprojectiles that were coated with cloned chloroplast DNA carrying the wild-type gene permanently restored the photosynthetic capacity of the algae. In most transformants of one of the mutants, a fragment with a 2.5-kilobase delection was restored to normal size by a homologous replacement event; in about 25 percent of the transformants the restored restriction fragment was 50 to 100 base pairs smaller or larger than that of wild type. About one-fourth of the transformants of this mutant contained unintegrated donor plasmid when first examined. This plasmid persisted in four different transformants after 65 cell generations of continuous liquid culture but was lost from all transformants maintained on plates of selective medium. The restored wild-type atpB gene remains in all transformants as an integral part of the chloroplast genome and is expressed and inherited normally.