ALTERATION OF CAENORHABDITIS-ELEGANS GENE-EXPRESSION BY TARGETED TRANSFORMATION

被引:19
作者
BROVERMAN, S [1 ]
MACMORRIS, M [1 ]
BLUMENTHAL, T [1 ]
机构
[1] INDIANA UNIV, DEPT BIOL, BLOOMINGTON, IN 47405 USA
关键词
GENE TARGETING; HOMOLOGOUS RECOMBINATION; PROMOTER FUNCTION; TRANSGENIC; NEMATODE;
D O I
10.1073/pnas.90.10.4359
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
We have produced strains carrying a synthetic fusion of parts of two vitellogenin genes, vit-2 and vit-6, integrated into the Caenorhabditis elegans genome. In most of the 63 transformant strains, the plasmid sequences are integrated at random locations in the genome. However, in two strains the transgene integrated by homologous recombination into the endogenous vit-2 gene. In both cases the reciprocal exchange between the chromosome and the injected circular plasmid containing a promoter deletion led to switching of the plasmid-borne promoter and the endogenous promoter, with a reduction in vit-2 expression. Thus in nematodes, transforming DNA can integrate by homologous recombination to result in partial inactivation of the chromosomal locus. The simplicity of the event and its reasonably high frequency suggest that gene targeting by homologous recombination should be considered as a method for directed inactivation of C. elegans genes.
引用
收藏
页码:4359 / 4363
页数:5
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