Diener D R, Curry A M, Johnson K A, Williams B D, Lefebvre P A, Kindle K L, Rosenbaum J L
Department of Biology, Yale University, New Haven, CT 06511.
Proc Natl Acad Sci U S A. 1990 Aug;87(15):5739-43. doi: 10.1073/pnas.87.15.5739.
The biflagellate alga Chlamydomonas has been used extensively in the genetic and biochemical analysis of flagellar assembly and motility. We have restored motility to a paralyzed-flagella mutant of Chlamydomonas by transforming with the corresponding wild-type gene. A nitrate reductase-deficient paralyzed-flagella strain, nit1-305 pf-14, carrying mutations in the genes for nitrate reductase and radial spoke protein 3, was transformed with wild-type copies of both genes. Two-thirds of the cells that survived nitrate selection also regained motility, indicating that they had been transformed with both the nitrate reductase and radial spoke protein 3 genes. Transformants typically contained multiple copies of both genes, genetically linked to each other, but not linked to the original mutant loci. Complementation of paralyzed-flagella mutants by transformation is a powerful tool for investigating flagellar assembly and function.
双鞭毛藻衣藻已被广泛用于鞭毛组装和运动的遗传与生化分析。我们通过用相应的野生型基因进行转化,使衣藻的一个鞭毛麻痹突变体恢复了运动能力。一个缺乏硝酸还原酶的鞭毛麻痹菌株nit1 - 305 pf - 14,其硝酸还原酶基因和辐条蛋白3基因发生了突变,用这两个基因的野生型拷贝进行了转化。在硝酸盐选择中存活下来的细胞中有三分之二也恢复了运动能力,这表明它们同时被硝酸还原酶基因和辐条蛋白3基因转化。转化体通常含有这两个基因的多个拷贝,它们在遗传上相互连锁,但与原始突变位点不连锁。通过转化对鞭毛麻痹突变体进行互补是研究鞭毛组装和功能的有力工具。