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Isolation of human transcribed sequences from human-rodent somatic cell hybrids
P Liu1, R Legerski, M J Siciliano
1Department of Molecular Genetics, University of Texas, M.D. Anderson Cancer Center, Houston 77030.
Abstract:
A method was developed for selectively isolating genes from localized regions of the human genome that are contained in interspecific hybrid cells. Complementary human DNA was prepared from a human-rodent somatic cell hybrid that contained less than 1% human DNA, by using consensus 5' intron splice sequences as primers. These primers would select immature, unspliced messenger RNA (still retaining species-specific repeat sequences) as templates. Screening a derived complementary DNA library for human repeat sequences resulted in the isolation of human clones at the anticipated frequency with characteristics expected of exons of transcribed human genes--single copy sequences that hybridized to discrete bands on Northern (RNA) blots.