EFFICIENT MASS TRANSFORMATION OF TETRAHYMENA-THERMOPHILA BY ELECTROPORATION OF CONJUGANTS

被引：107

作者：

GAERTIG, J

GOROVSKY, MA

机构：

[1] Department of Biology, University of Rochester, Rochester

来源：

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA | 1992年 / 89卷 / 19期

关键词：

CILIATE; RIBOSOMAL PROTEIN L29 GENE; RIBOSOMAL-RNA GENE; CYCLOHEXIMIDE; PAROMOMYCIN;

D O I：

10.1073/pnas.89.19.9196

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

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 mug 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.

引用

页码：9196 / 9200

页数：5

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