TY - JOUR
T1 - Direct selection of expressed sequences within a 1-Mb region flanking BRCA1 on human chromosome 17q21
AU - Osborne-Lawrence, Sherri
AU - Welcsh, Piri L.
AU - Spillman, Monique
AU - Chandrasekharappa, Settara C.
AU - Gallardo, Teresa D.
AU - Lovett, Michael
AU - Bowcock, Anne M.
N1 - Funding Information:
This study was partly supported by NIH Grant CA60650 (A.B.), NIH Grants HG00368 and HG00882 to M.L., and a Susan G. Komen Fellowship to Piri Welcsh. Monique Spillman was supported in part by a grant from the Perot Family Foundation. Technical assistance was provided by Corky Dunn and To Thai. We thank Greg Clines for preparation of the RNU2 YACs and Dr. Bento Soares (Columbia University) for the normalized infant brain cDNA library.
PY - 1995/1/1
Y1 - 1995/1/1
N2 - Direct selection of genes within the interval of chromosome 17q21 containing BRCA1 was performed. YAC and cosmid contigs spanning the BRCA1 region were used to select cDNA clones from pools of cDNAs derived from human placenta, HeLa cells, activated T cells, and fetal head. A minimum set of 48 fragments of nonoverlapping cDNAs that unequivocally mapped within a 1-Mb region was identified, although it is not yet known how many of these are derived from the same transcript. DNA sequence analyses revealed that 4 of these cDNAs were derived from known genes (EDH17B2, glucose-6-phosphatase, IAI.3B, and E1AF), 1 is a member of a previously described gene family (HMG-17), and 7 share substantial identity with previously described genes from human or other species. The remainder showed no significant homology to known genes. Limited PCR-based expression profiles of a set of 13 of the genes were performed, and all gave positive results with at least some cDNA sources supporting the contention that they truly represent transcribed sequences. A comparison between genes obtained from this region by direct selection with those obtained by direct screening or exon trapping (see accompanying papers, this issue) revealed that over 90% of the genes identified by exon trapping were represented in the selected material and that at least two additional genes that appear to represent low abundance transcripts with restricted expression profiles were identified by selection but not by other means.
AB - Direct selection of genes within the interval of chromosome 17q21 containing BRCA1 was performed. YAC and cosmid contigs spanning the BRCA1 region were used to select cDNA clones from pools of cDNAs derived from human placenta, HeLa cells, activated T cells, and fetal head. A minimum set of 48 fragments of nonoverlapping cDNAs that unequivocally mapped within a 1-Mb region was identified, although it is not yet known how many of these are derived from the same transcript. DNA sequence analyses revealed that 4 of these cDNAs were derived from known genes (EDH17B2, glucose-6-phosphatase, IAI.3B, and E1AF), 1 is a member of a previously described gene family (HMG-17), and 7 share substantial identity with previously described genes from human or other species. The remainder showed no significant homology to known genes. Limited PCR-based expression profiles of a set of 13 of the genes were performed, and all gave positive results with at least some cDNA sources supporting the contention that they truly represent transcribed sequences. A comparison between genes obtained from this region by direct selection with those obtained by direct screening or exon trapping (see accompanying papers, this issue) revealed that over 90% of the genes identified by exon trapping were represented in the selected material and that at least two additional genes that appear to represent low abundance transcripts with restricted expression profiles were identified by selection but not by other means.
UR - http://www.scopus.com/inward/record.url?scp=0028963322&partnerID=8YFLogxK
U2 - 10.1016/0888-7543(95)80132-6
DO - 10.1016/0888-7543(95)80132-6
M3 - Article
C2 - 7774925
AN - SCOPUS:0028963322
SN - 0888-7543
VL - 25
SP - 248
EP - 255
JO - Genomics
JF - Genomics
IS - 1
ER -