Direct cloning of human transcripts with HnRNA from hybrid cell lines.
A library of human-derived complementary DNA from a human-hamster hybrid cell line containing the Xq24-qter region has been constructed. Complementary DNA synthesis was primed from heterogeneous nuclear (hn) RNA by oligonucleotides derived from conserved regions of human Alu repeats. At least 80% of these cloned sequences were of human origin, providing an enrichment of at least two orders of magnitude. Two clones, one containing a fragment of the primary transcript of the human hypoxanthine-guanine phosphoribosyltransferase (HPRT) gene at Xq26 and another recognizing a family of human genes mapping to two regions of Xq24-qter, were characterized. Additional hncDNA clones mapped to a variety of sites in the Xq24-qter region, demonstrating the isolation of many transcriptionally active loci. These clones provide probes for identification of genetic loci on the terminal region of the X chromosome long arm, which is the location of a number of inherited disorders.
Corbo
L
L
Institute for Molecular Genetics, Baylor College of Medicine, Houston, TX 77030.
Maley
J A
JA
Nelson
D L
DL
Caskey
C T
CT
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002874:Chromosome Mapping; D003001:Cloning, Molecular / Q000379:methods; D006224:Cricetinae; D004277:DNA, Single-Stranded / Q000235:genetics / Q000302:isolation & purification; D015246:Deoxyribonuclease EcoRI; D006801:Humans; D006822:Hybrid Cells / Q000166:cytology; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes; D012332:RNA, Heterogeneous Nuclear / Q000235:genetics*; D015183:Restriction Mapping; D014158:Transcription, Genetic; D014960:X Chromosome
D004277:DNA, Single-Stranded; D015345:Oligonucleotide Probes; D012332:RNA, Heterogeneous Nuclear; D015246:Deoxyribonuclease EcoRI