Gaertig J, Gorovsky M A
Department of Biology, University of Rochester, NY 14627.
Proc Natl Acad Sci U S A. 1992 Oct 1;89(19):9196-200. doi: 10.1073/pnas.89.19.9196.
Conjugating cells of the ciliate Tetrahymena thermophila were electroporated in the presence of plasmid DNA containing a paromomycin-resistant ribosomal RNA gene (rDNA). Cells were selected with paromomycin following 12-24 hr of growth on nonselective medium. Resistant cells appeared after 2-3 days. Processing vectors containing the micronuclear rDNA and somatic vectors containing the macronuclear gene transformed the cells, with the former yielding frequencies up to 900 transformants per microgram of plasmid DNA. A ribosomal protein gene (rpL29) conferring cycloheximide resistance also transformed conjugating cells. The transformation efficiency of the plasmid containing only the rpL29 gene was increased by insertion of an rDNA replication origin and by cotransformation and preselection with an rDNA vector. These results indicate that electroporation can be used for the production of large numbers of transformed Tetrahymena.
在含有对巴龙霉素耐药的核糖体RNA基因（rDNA）的质粒DNA存在的情况下，对嗜热栖热四膜虫的接合细胞进行电穿孔处理。细胞在非选择性培养基上生长12 - 24小时后，用巴龙霉素进行筛选。2 - 3天后出现抗性细胞。含有微核rDNA的加工载体和含有大核基因的体细胞载体转化了细胞，前者每微克质粒DNA产生的转化体频率高达900个。赋予环己酰亚胺抗性的核糖体蛋白基因（rpL29）也转化了接合细胞。通过插入rDNA复制起点以及与rDNA载体共转化和预筛选，仅含有rpL29基因的质粒的转化效率得以提高。这些结果表明电穿孔可用于大量生产转化的四膜虫。
