Yeast artificial chromosomes containing human Xq24-Xq28 DNA: library construction and representation of probe sequences - PubMed (original) (raw)
Yeast artificial chromosomes containing human Xq24-Xq28 DNA: library construction and representation of probe sequences
F E Abidi et al. Genomics. 1990 Jul.
Abstract
A library of yeast artificial chromosomes (YACs) with human DNA inserts has been assembled from a human/hamster somatic cell hybrid containing Xq24-Xqter human DNA. Screening of the agar-embedded transformants for human DNA used a manifold of 3000 stainless-steel pins to transfer colonies onto the surface of media. This facilitated the recovery of the 1 in 300 clones that contained a human DNA insert (the remainder had hamster DNA and were discarded). The library described here consists of about two genomic equivalents (102 Mb) of human DNA in 467 clones: 167 were generated by EcoRI partial digestion and contain 25.5 Mb of human DNA; 252 used partial digestion with TaqI and cover 64.2 Mb; and 48 were from sheared DNA inserts and cover 11.7 Mb. Clones were screened by hybridization with 70 probes previously assigned to Xq24-Xq28. Eleven probes did not hybridize to any YACs in the library, and 16 probes hybridized to one YAC each, 23 to two, 13 to three, and 7 to four. Also, individual YACs large enough to detect features like the clustering of polymorphic sequences in subregions of Xq24-Xqter have been obtained. For example, XY58 contained five probe sequences previously independently isolated. The overall yield of YACs containing probe sequences was indistinguishable from Poisson statistical expectations for random cloning (P = 0.9). Thus, YAC libraries such as the one described here can include most, if not all, of the sequences in the source DNA from which the library is derived. These results support the possibility that YACs may provide a reliable bridge between linkage studies and conventional recombinant DNA analyses in mapping of the human genome.
Similar articles
- Human Xq24-Xq28: approaches to mapping with yeast artificial chromosomes.
Wada M, Little RD, Abidi F, Porta G, Labella T, Cooper T, Della Valle G, D'Urso M, Schlessinger D. Wada M, et al. Am J Hum Genet. 1990 Jan;46(1):95-106. Am J Hum Genet. 1990. PMID: 2294758 Free PMC article. - In situ hybridization to cytogenetic bands of yeast artificial chromosomes covering 50% of human Xq24-Xq28 DNA.
Montanaro V, Casamassimi A, D'Urso M, Yoon JY, Freije W, Schlessinger D, Muenke M, Nussbaum RL, Saccone S, Maugeri S, et al. Montanaro V, et al. Am J Hum Genet. 1991 Feb;48(2):183-94. Am J Hum Genet. 1991. PMID: 1990831 Free PMC article. - Yeast artificial chromosomes with 200- to 800-kilobase inserts of human DNA containing HLA, V kappa, 5S, and Xq24-Xq28 sequences.
Little RD, Porta G, Carle GF, Schlessinger D, D'Urso M. Little RD, et al. Proc Natl Acad Sci U S A. 1989 Mar;86(5):1598-602. doi: 10.1073/pnas.86.5.1598. Proc Natl Acad Sci U S A. 1989. PMID: 2537978 Free PMC article. - Yeast artificial chromosome-based genome mapping: some lessons from Xq24-q28.
Schlessinger D, Little RD, Freije D, Abidi F, Zucchi I, Porta G, Pilia G, Nagaraja R, Johnson SK, Yoon JY, Srivastava A, Kere J, Palmieri G, Ciccodicola A, Montanaro V, Romano G, Casamassimi A, D'Urso M. Schlessinger D, et al. Genomics. 1991 Dec;11(4):783-93. doi: 10.1016/0888-7543(91)90001-u. Genomics. 1991. PMID: 1783389 Review. - Yeast artificial chromosomes (YACs) and the analysis of complex genomes.
Anand R. Anand R. Trends Biotechnol. 1992 Jan-Feb;10(1-2):35-40. doi: 10.1016/0167-7799(92)90165-r. Trends Biotechnol. 1992. PMID: 1367930 Review.
Cited by
- Specific cloning of human DNA as yeast artificial chromosomes by transformation-associated recombination.
Larionov V, Kouprina N, Graves J, Chen XN, Korenberg JR, Resnick MA. Larionov V, et al. Proc Natl Acad Sci U S A. 1996 Jan 9;93(1):491-6. doi: 10.1073/pnas.93.1.491. Proc Natl Acad Sci U S A. 1996. PMID: 8552668 Free PMC article. - An integrated approach for identifying and mapping human genes.
Das Gupta R, Morrow B, Marondel I, Parimoo S, Goei VL, Gruen J, Weissman S, Skoultchi A, Kucherlapati R. Das Gupta R, et al. Proc Natl Acad Sci U S A. 1993 May 15;90(10):4364-8. doi: 10.1073/pnas.90.10.4364. Proc Natl Acad Sci U S A. 1993. PMID: 8506274 Free PMC article. - Low-frequency chimeric yeast artificial chromosome libraries from flow-sorted human chromosomes 16 and 21.
McCormick MK, Campbell E, Deaven L, Moyzis R. McCormick MK, et al. Proc Natl Acad Sci U S A. 1993 Feb 1;90(3):1063-7. doi: 10.1073/pnas.90.3.1063. Proc Natl Acad Sci U S A. 1993. PMID: 8430075 Free PMC article. - Initiation and termination of DNA replication in human rRNA genes.
Little RD, Platt TH, Schildkraut CL. Little RD, et al. Mol Cell Biol. 1993 Oct;13(10):6600-13. doi: 10.1128/mcb.13.10.6600-6613.1993. Mol Cell Biol. 1993. PMID: 8413256 Free PMC article. - Chimeric YACs were generated at unreduced rates in conditions that suppress coligation.
Wada M, Abe K, Okumura K, Taguchi H, Kohno K, Imamoto F, Schlessinger D, Kuwano M. Wada M, et al. Nucleic Acids Res. 1994 May 11;22(9):1651-4. doi: 10.1093/nar/22.9.1651. Nucleic Acids Res. 1994. PMID: 8202367 Free PMC article.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Miscellaneous