Chromosome assignment of human brain expressed sequence tags (ESTs) by analyzing fluorescently labeled PCR products from hybrid cell panels.
Sixty-three human brain cDNA sequences were newly assigned to individual human chromosomes. Ten of these were subregionally localized, and one was also mapped in the mouse genome. Four previously reported assignments were refined. PCR primers were designed from expressed sequence tags (ESTs) and tested for specific amplification from human genomic DNA. DNA was then amplified, often in multiplexed PCR reactions, using DNA from somatic cell hybrid mapping panels as templates. The amplification products were identified using an automated fluorescence detection system. Chromosomal assignments were made by discordancy analysis. Thirteen newly localized cDNAs exhibited homology to previously reported sequences. EST01471 was shown to correspond to human microtubule-associated protein 1B (MAP1B), confirming the previous mapping of this gene to human chromosome 5. Other genes tentatively assigned to chromosomes based on these results were a component of the signal peptide receptor of the endoplasmic reticulum (EST00745) and a cyclic AMP-regulated phosphoprotein (EST01041) on chromosome 1, a protein phosphatase 2A 55-kDa regulatory subunit (EST01650) on chromosome 4, an NAD(P) transhydrogenase (EST01744) on chromosome 5, ribosomal proteins L1a or L1b (EST01627) and L18a (EST01583), a brain transcription factor (BF-1, EST00795) on chromosome 14, a milk fat globule membrane-related protein (EST01678) on chromosome 15, a putative peptide initiation factor (EST00675) on chromosome 17, thiosulfate sulfurtransferase (TST) on chromosome 22, and moesin (MSN) (EST00896) and a human equivalent of rat spot 14 (S14) (EST00887) on Xp11-->cen and Xpter-->p21.3, respectively.
Durkin
A S
AS
Department of Molecular Biology, American Type Culture Collection, Rockville, MD.
Nierman
W C
WC
Zoghbi
H
H
Jones
C
C
Kozak
C A
CA
Maglott
D R
DR
D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000378:metabolism*; D002874:Chromosome Mapping; D006224:Cricetinae; D004247:DNA; D018076:DNA, Complementary / Q000235:genetics*; D005456:Fluorescent Dyes; D015870:Gene Expression; D006801:Humans; D006822:Hybrid Cells; D007256:Information Systems; D051379:Mice; D008869:Microtubule-Associated Proteins; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction
D018076:DNA, Complementary; D005456:Fluorescent Dyes; D008869:Microtubule-Associated Proteins; C085284:microtubule-associated protein 1B; D004247:DNA