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Proc Natl Acad Sci U S A. Nov 1, 1991; 88(21): 9628–9632.

Direct selection: a method for the isolation of cDNAs encoded by large genomic regions.


We have developed a strategy for the rapid enrichment and identification of cDNAs encoded by large genomic regions. The basis of this "direct selection" scheme is the hybridization of an entire library of cDNAs to an immobilized genomic clone. Nonspecific hybrids are eliminated and selected cDNAs are eluted. These molecules are then amplified and are either cloned or subjected to further selection/amplification cycles. This scheme was tested using a 550-kilobase yeast artificial chromosome clone that contains the EPO gene. Using this clone and a fetal kidney cDNA library, we have achieved a 1000-fold enrichment of EPO cDNAs in one cycle of enrichment. More significantly, we have further investigated one of the "anonymous" cDNAs that was selectively enriched. We confirmed that this cDNA was encoded by the yeast artificial chromosome. Its frequency in the starting library was 1 in 1 x 10(5) cDNAs and after selection comprised 2% of the selected library. DNA sequence analysis of this cDNA and of the yeast artificial chromosome clone revealed that this gene encodes the beta 2 subunit of the human guanine nucleotide-binding regulatory proteins. Restriction mapping and hybridization data position this gene (GNB2) to within 30-70 kilobases of the EPO gene. The selective isolation and mapping of GNB2 confirms the feasibility of this direct selection strategy and suggests that it will be useful for the rapid isolation of cDNAs, including disease-related genes, across extensive portions of the human genome.

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  • Lindsay S, Bird AP. Use of restriction enzymes to detect potential gene sequences in mammalian DNA. Nature. 327(6120):336–338. [PubMed]
  • Lavia P, Macleod D, Bird A. Coincident start sites for divergent transcripts at a randomly selected CpG-rich island of mouse. EMBO J. 1987 Sep;6(9):2773–2779. [PMC free article] [PubMed]
  • Pontarotti P, Chimini G, Nguyen C, Boretto J, Jordan BR. CpG islands and HTF islands in the HLA class I region: investigation of the methylation status of class I genes leads to precise physical mapping of the HLA-B and -C genes. Nucleic Acids Res. 1988 Jul 25;16(14B):6767–6778. [PMC free article] [PubMed]
  • Abe K, Wei JF, Wei FS, Hsu YC, Uehara H, Artzt K, Bennett D. Searching for coding sequences in the mammalian genome: the H-2K region of the mouse MHC is replete with genes expressed in embryos. EMBO J. 1988 Nov;7(11):3441–3449. [PMC free article] [PubMed]
  • Rommens JM, Iannuzzi MC, Kerem B, Drumm ML, Melmer G, Dean M, Rozmahel R, Cole JL, Kennedy D, Hidaka N, et al. Identification of the cystic fibrosis gene: chromosome walking and jumping. Science. 1989 Sep 8;245(4922):1059–1065. [PubMed]
  • Yokoi T, Lovett M, Cheng ZY, Epstein CJ. Isolation of transcribed DNA sequences from chromosome 21 using mouse fetal cDNA. Hum Genet. 1986 Oct;74(2):137–142. [PubMed]
  • Hochgeschwender U, Sutcliffe JG, Brennan MB. Construction and screening of a genomic library specific for mouse chromosome 16. Proc Natl Acad Sci U S A. 1989 Nov;86(21):8482–8486. [PMC free article] [PubMed]
  • Saiki RK, Gelfand DH, Stoffel S, Scharf SJ, Higuchi R, Horn GT, Mullis KB, Erlich HA. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. [PubMed]
  • Liu P, Legerski R, Siciliano MJ. Isolation of human transcribed sequences from human-rodent somatic cell hybrids. Science. 1989 Nov 10;246(4931):813–815. [PubMed]
  • Duyk GM, Kim SW, Myers RM, Cox DR. Exon trapping: a genetic screen to identify candidate transcribed sequences in cloned mammalian genomic DNA. Proc Natl Acad Sci U S A. 1990 Nov;87(22):8995–8999. [PMC free article] [PubMed]
  • Buckler AJ, Chang DD, Graw SL, Brook JD, Haber DA, Sharp PA, Housman DE. Exon amplification: a strategy to isolate mammalian genes based on RNA splicing. Proc Natl Acad Sci U S A. 1991 May 1;88(9):4005–4009. [PMC free article] [PubMed]
  • Hochgeschwender U, Brennan MB. Identifying genes within the genome: new ways for finding the needle in a haystack. Bioessays. 1991 Mar;13(3):139–144. [PubMed]
  • Burke DT, Carle GF, Olson MV. Cloning of large segments of exogenous DNA into yeast by means of artificial chromosome vectors. Science. 1987 May 15;236(4803):806–812. [PubMed]
  • Wallace MR, Marchuk DA, Andersen LB, Letcher R, Odeh HM, Saulino AM, Fountain JW, Brereton A, Nicholson J, Mitchell AL, et al. Type 1 neurofibromatosis gene: identification of a large transcript disrupted in three NF1 patients. Science. 1990 Jul 13;249(4965):181–186. [PubMed]
  • Elvin P, Slynn G, Black D, Graham A, Butler R, Riley J, Anand R, Markham AF. Isolation of cDNA clones using yeast artificial chromosome probes. Nucleic Acids Res. 1990 Jul 11;18(13):3913–3917. [PMC free article] [PubMed]
  • Sealey PG, Whittaker PA, Southern EM. Removal of repeated sequences from hybridisation probes. Nucleic Acids Res. 1985 Mar 25;13(6):1905–1922. [PMC free article] [PubMed]
  • Moyzis RK, Torney DC, Meyne J, Buckingham JM, Wu JR, Burks C, Sirotkin KM, Goad WB. The distribution of interspersed repetitive DNA sequences in the human genome. Genomics. 1989 Apr;4(3):273–289. [PubMed]
  • Burke DT, Olson MV. Preparation of clone libraries in yeast artificial-chromosome vectors. Methods Enzymol. 1991;194:251–270. [PubMed]
  • Church GM, Gilbert W. Genomic sequencing. Proc Natl Acad Sci U S A. 1984 Apr;81(7):1991–1995. [PMC free article] [PubMed]
  • Patanjali SR, Parimoo S, Weissman SM. Construction of a uniform-abundance (normalized) cDNA library. Proc Natl Acad Sci U S A. 1991 Mar 1;88(5):1943–1947. [PMC free article] [PubMed]
  • Crampton JM, Davies KE, Knapp TF. The occurrence of families of repetitive sequences in a library of cloned cDNA from human lymphocytes. Nucleic Acids Res. 1981 Aug 11;9(15):3821–3834. [PMC free article] [PubMed]
  • Watkins PC, Eddy R, Hoffman N, Stanislovitis P, Beck AK, Galli J, Vellucci V, Gusella JF, Shows TB. Regional assignment of the erythropoietin gene to human chromosome region 7pter----q22. Cytogenet Cell Genet. 1986;42(4):214–218. [PubMed]
  • Law ML, Cai GY, Lin FK, Wei Q, Huang SZ, Hartz JH, Morse H, Lin CH, Jones C, Kao FT. Chromosomal assignment of the human erythropoietin gene and its DNA polymorphism. Proc Natl Acad Sci U S A. 1986 Sep;83(18):6920–6924. [PMC free article] [PubMed]
  • Fong HK, Amatruda TT, 3rd, Birren BW, Simon MI. Distinct forms of the beta subunit of GTP-binding regulatory proteins identified by molecular cloning. Proc Natl Acad Sci U S A. 1987 Jun;84(11):3792–3796. [PMC free article] [PubMed]
  • Blatt C, Eversole-Cire P, Cohn VH, Zollman S, Fournier RE, Mohandas LT, Nesbitt M, Lugo T, Jones DT, Reed RR, et al. Chromosomal localization of genes encoding guanine nucleotide-binding protein subunits in mouse and human. Proc Natl Acad Sci U S A. 1988 Oct;85(20):7642–7646. [PMC free article] [PubMed]
  • Bray P, Carter A, Guo V, Puckett C, Kamholz J, Spiegel A, Nirenberg M. Human cDNA clones for an alpha subunit of Gi signal-transduction protein. Proc Natl Acad Sci U S A. 1987 Aug;84(15):5115–5119. [PMC free article] [PubMed]
  • Bloch DB, Bloch KD, Iannuzzi M, Collins FS, Neer EJ, Seidman JG, Morton CC. The gene for the alpha i1 subunit of human guanine nucleotide binding protein maps near the cystic fibrosis locus. Am J Hum Genet. 1988 Jun;42(6):884–888. [PMC free article] [PubMed]
  • Bickmore WA, Sumner AT. Mammalian chromosome banding--an expression of genome organization. Trends Genet. 1989 May;5(5):144–148. [PubMed]

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