© 1992 Oxford University Press
RESEARCH-ARTICLE |
A strategy for the selection of transcribed sequences in the Xq28 region
1Imperial Cancer Research Fund, 44 Uncoln's Inn Fields London WC2A 3PX, UK Deutsches Krebsforschungszentrum, Im Neuenheimer Feld 280 6900 Heidelberg, Germany
*To whom correspondence should be addressed
Received May 29, 1992; Revised February 25, 1989; Accepted June 25, 1992
As an essential step towards an exhaustive analysis of the coding potential of large regions of the genome, we have developed a protocol allowing the rapid isolation of transcripts defmed by overlapping clone libraries. The method is based on the hybridisation of cDNA inserts, which had been amplified by PCR from cDNA libraries, to biotiiiylated DNA from cosmids or cosmid pools. Nonspecific hybrids are then removed, the selected cDNAs are eluted and reamplifled by PCR. Using a cosmid containing part of the FMR-1 gene as test, we were able to demonstrate an eighty thousand fold enrichment of cDNAs for this gene after two rounds of selection-amplification. The technique was applied to the analysis of transcripts from two cosmid contigs, together encompassing a region of 900 kb in Xq28. These experiments have thus far resulted in the identification of 81 cDNA clones, of which 54 clones were mapped back to the cosmid contigs. Of the 54 clones placed on the contig maps, 12 cDNA clones can be shown to belong to two genes which have been previously reported (L1CAM and QM).
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  J. L. Joyal, J. Jiang, and D. I. Israel Use of PCR to Prepare a Double-Stranded DNA Library Encoding Random Peptides CSH Protocols, May 1, 2006; 2006(14): pdb.prot4135 - pdb.prot4135. [Full Text]
|
|
|
|
 |
|
 B. Gust, K. Spatz, A. Spychaj, and M. Redenbach Region-Specific Transcriptional Activity in the Genome of Streptomyces coelicolor A3(2) Appl. Envir. Microbiol., August 1, 2001; 67(8): 3598 - 3602. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 N. Tripodis, S. Palmer, S. Phillips, S. Milne, S. Beck, and J. Ragoussis Construction of a High-Resolution 2.5-Mb Transcript Map of the Human 6p21.2-6p21.3 Region Immediately Centromeric of the Major Histocompatibility Complex Genome Res., April 1, 2000; 10(4): 454 - 472. [Abstract] [Full Text]
|
|
|
|
 |
|
 J. Gilley and M. Fried Extensive gene order differences within regions of conserved synteny between the Fugu and human genomes: implications for chromosomal evolution and the cloning of disease genes Hum. Mol. Genet., July 1, 1999; 8(7): 1313 - 1320. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Hildmann, X. Kong, J. O'Brien, L. Riesselman, H.-M. Christensen, E. Dagand, H. Lehrach, and M.-L. Yaspo A Contiguous 3-Mb Sequence-Ready Map in the S3-MX Region on 21q22.2 Based on High- Throughput Nonisotopic Library Screenings Genome Res., April 1, 1999; 9(4): 360 - 372. [Abstract] [Full Text]
|
|
|
|
|
|
J. M. McKie, R. B. Wadey, H. F. Sutherland, C. L. Taylor, and P. J. Scambler Direct Selection of Conserved cDNAs from the DiGeorge Critical Region: Isolation of a Novel CDC45-Like Gene Genome Res., August 1, 1998; 8(8): 834 - 841. [Abstract] [Full Text]
|
|
|
|
|
|
N S Heiss, U C Rogner, P Kioschis, B Korn, and A Poustka Transcription mapping in a 700-kb region around the DXS52 locus in Xq28: isolation of six novel transcripts and a novel ATPase isoform (hPMCA5). Genome Res., June 1, 1996; 6(6): 478 - 491. [Abstract] [PDF]
|
|
|
|
|
|
D P Mortlock, M R Nelson, and J W Innis An efficient method for isolating putative promoters and 5'-transcribed sequences from large genomic clones. Genome Res., April 1, 1996; 6(4): 327 - 335. [Abstract] [PDF]
|
|
|
|
 |
|
 S. Rouquier, J. B. Lowe, R. J. Kelly, A. L. Fertitta, G. G. Lennon, and D. Giorgi Molecular Cloning of a Human Genomic Region Containing the H Blood Group alpha(1,2)Fucosyltransferase Gene and Two H Locus-related DNA Restriction Fragments J. Biol. Chem., March 3, 1995; 270(9): 4632 - 4639. [Abstract] [Full Text] [PDF]
|
|