Creation of low-copy integrated transgenic lines in Caenorhabditis elegans.

摘要: In Caenorhabditis elegans, transgenic lines are typically created by injecting DNA into the hermaphrodite germline to form multicopy extrachromosomal arrays. This technique is a reliable means of expressing transgenes in C. but its use has limitations. Because arrays semistable, only fraction animals array line transformed. addition, because can contain hundreds copies transforming DNA, may be overexpressed, misexpressed, or silenced. We have developed an alternative method for elegans transformation, using microparticle bombardment, that produces single- and low-copy chromosomal insertions. Using this method, we find it possible create integrated reproducibly express GFP reporter constructs without variations expression level pattern frequently exhibited lines. also used germline, where conventional fail due silencing.