Effect of cis-located human satellite DNA on the electroporation efficiency of neo and HSV-1 tk containing plasmids.

Highly repetitive satellite DNAs comprise a significant portion of higher eukaryotic genomes and have been implicated in a variety of chromosome processes, such as centromere structure and function, that are related to their presence in heterochromatin. In addition, heterochromatin can induce metastable expression of adjacent genes. However, the role of highly repetitive satellite DNAs in these effects remains to be elucidated. In an effort to address this question, plasmids containing a human 1797-bp EcoRI satellite II DNA, plus the neo and the HSV-1 tk genes, were electroporated into a TK-/NEO- human cell line. The presence of the satellite DNA sequences within the electroporated plasmids was found to interfere with the generation of stable TK+, but not NEO+, transfectants depending on the location and/or orientation of the cloned satellite DNA.