Boynton J E, Gillham N W, Harris E H, Hosler J P, Johnson A M, Jones A R, Randolph-Anderson B L, Robertson D, Klein T M, Shark K B
Department of Botany, Duke University, Durham, NC 27706.
Science. 1988 Jun 10;240(4858):1534-8. doi: 10.1126/science.2897716.
Bombardment of three mutants of the chloroplast 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 deletion 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.
用包被有携带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，永久性地恢复了藻类的光合能力。在其中一个突变体的大多数转化体中，一个有2.5千碱基缺失的片段通过同源替换事件恢复到正常大小；在约25%的转化体中，恢复的限制片段比野生型的小或大50至100个碱基对。首次检测时，该突变体约四分之一的转化体含有未整合的供体质粒。在连续液体培养65个细胞世代后，该质粒在四个不同的转化体中持续存在，但在选择性培养基平板上培养的所有转化体中均丢失。恢复的野生型atpB基因作为叶绿体基因组的一个组成部分保留在所有转化体中，并正常表达和遗传。
