Isolation of human transcribed sequences from human-rodent somatic cell hybrids.
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.
Liu
P
P
Department of Molecular Genetics, University of Texas, M.D. Anderson Cancer Center, Houston 77030.
Legerski
R
R
Siciliano
M J
MJ
D000818:Animals; D015152:Blotting, Northern; D015139:Blotting, Southern; D002874:Chromosome Mapping; D002888:Chromosomes, Human, Pair 19; D003001:Cloning, Molecular; D006224:Cricetinae; D004247:DNA / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D006822:Hybrid Cells; D007438:Introns; D009693:Nucleic Acid Hybridization; D012313:RNA / Q000235:genetics; D012091:Repetitive Sequences, Nucleic Acid; D015183:Restriction Mapping; D013698:Templates, Genetic
D012313:RNA; D004247:DNA