Recombination‐Based Assay (RBA) for Screening Bacteriophage Lambda Libraries

Show simple item record Kurnit, David M. 2020-01-13T15:21:36Z 2020-01-13T15:21:36Z 1994-07
dc.identifier.citation Kurnit, David M. (1994). "Recombination‐Based Assay (RBA) for Screening Bacteriophage Lambda Libraries." Current Protocols in Molecular Biology 27(1): 6.12.1-6.12.12.
dc.identifier.issn 1934-3639
dc.identifier.issn 1934-3647
dc.description.abstract The recombination‐based assay represents a convenient way to screen a complex library constructed in bacteriophage l for homology to a given sequence cloned into a specially designed plasmid. The technique serves to screen a bacteriophage library rapidly and efficiently with a sequence cloned into a plasmid; counterselection then yields the gene product of interest with its plasmid carrier deleted. Because 106 to 107 plaque‐forming units (pfu) may be screened using several petri dishes, and the homology for crossing‐over need only be greater than 25 bp, the RBA represents an efficient way to screen complex l libraries rapidly for homology to a given sequence. In this procedure, a l library is screened using a specially designed plasmid carrying the desired target sequence. Recombinants arising from cross‐over events between the plasmid and a bacteriophage carrying a corresponding region of homology are selected by their ability to grow on strain DM21. Growth of l on DM21 requires the presence of an allele encoded on the plasmid to suppress an amber mutation in the host strain that prevents l propagation. Recovery of the original phage carrying the target sequence requires a reversal of the homologous recombination event. This reversal occurs spontaneously, and is detected by PCR amplification using primers that flank the cloning site in the l vector.
dc.publisher John Wiley & Sons
dc.title Recombination‐Based Assay (RBA) for Screening Bacteriophage Lambda Libraries
dc.type Article
dc.rights.robots IndexNoFollow
dc.subject.hlbsecondlevel Molecular, Cellular and Developmental Biology
dc.subject.hlbsecondlevel Genetics
dc.subject.hlbtoplevel Health Sciences
dc.subject.hlbtoplevel Science
dc.description.peerreviewed Peer Reviewed
dc.identifier.doi 10.1002/0471142727.mb0612s27
dc.identifier.source Current Protocols in Molecular Biology
dc.identifier.citedreference Rubin, C.M., Houck, C.M., Deininger, P.L., and Schmid, C.W. 1980. Partial nucleotide sequence of the 300 nucleotide interspersed repeated human DNA sequences. Nature 284: 372 ‐ 374.
dc.identifier.citedreference Saiki, R.K., Scharf, S., Faloona, F., Mullis, K.B., Horn, G., Erlich, H.A., and Arnheim, N. 1985. Enzymatic amplification of β‐globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. Science 230: 1350 ‐ 1354.
dc.identifier.citedreference Young, R.A. and Davis, R.W. 1983. Efficient isolation of genes by using antibody probes. Proc. Natl. Acad. Sci. U.S.A. 80: 1194 ‐ 1198.
dc.identifier.citedreference Yanisch‐Perron, C., Vieira, J., and Messing, J. 1985. Improved M13 phage cloning vectors and host strains: Nucleotide sequences of the M13mp18 and pUC19 vectors. Gene 33: 103 ‐ 119.
dc.identifier.citedreference Watt, V.M., Ingles, C.J., Urdea, M.S., and Rutter, W.J. 1985. Homology requirements for recombination in Escherichia coli. Proc. Natl. Acad. Sci. U.S.A. 82: 4768 ‐ 4772.
dc.identifier.citedreference Stewart, G.D., Hauser, M.A., Kang, H., McCann, D.P., Osemlak, M.M., Kurnit, D.M., and Hanzlik, A.J. 1991. Plasmids for recombination‐based screening. Gene 106: 97 ‐ 101.
dc.identifier.citedreference Short, J.M., Fernandez, J.M., Sorge, J.A., and Huse, W.D. 1988. λ ZAP: A bacteriophage λ expression vector with in vivo excision properties. Nucl. Acids Res. 16: 7583 ‐ 7599.
dc.identifier.citedreference Shen, P. and Huang, H.V. 1989. Effect of base pair mismatches on recombination via the recBCD pathway. Mol. Gen. Genet. 218: 358 ‐ 360.
dc.identifier.citedreference Marvo, S.L., King, S.R., and Jaskunas, S.R. 1983. Role of short regions of homology in intermolecular illegitimate recombination events. Proc. Natl. Acad. Sci. U.S.A. 80: 2452 ‐ 2456.
dc.identifier.citedreference Neve, R.L. and Kurnit, D.M. 1983. Comparison of sequence repetitiveness of human cDNA and genomic DNA using the miniplasmid vector piVX. Gene 23: 355 ‐ 367.
dc.identifier.citedreference Shen, P. and Huang, H.V. 1986. Homologous recombination in Escherichia coli: Dependence on substrate length and homology. Genetics 112: 441 ‐ 457.
dc.identifier.citedreference Neve, R.L., Bruns, G.A.P., Dryja, T.P., and Kurnit, D.M. 1983. Retrieval of human DNA from rodent‐human genomic libraries by a recombination process. Gene 23: 343 ‐ 354.
dc.identifier.citedreference Seed, B. 1983. Purification of genomic sequences from bacteriophage libraries by recombination and selection in vivo. Nucl. Acids Res. 11: 2427 ‐ 2445.
dc.identifier.citedreference Bolivar, F., Rodriguez, R., Green, P.J., Betlach, M., Heyneker, H.L., Boyer, H.W., Crosa, J., and Falkow, S. 1977. Construction and characterization of new cloning vehicles. Gene 2: 95 ‐ 113.
dc.identifier.citedreference Buckler, A.J., Chang, D.D., Graw, S.L., Brook, J.D., Haber, D.A., Sharp, P.A., and Housman, D.E. 1991. Exon amplificaton: A strategy to isolate mammalian genes based on RNA splicing. Proc. Natl. Acad. Sci. U.S.A. 88: 4005 ‐ 4009.
dc.identifier.citedreference Casadaban, M.J. and Cohen, S.N. 1980. Analysis of gene control signals by DNA fusion and cloning in Escherichia coli. J. Mol. Biol. 138: 179 ‐ 207.
dc.identifier.citedreference Duyk, G.M., Kim, S., Myers, R.M., and Cox, D.R. 1990. Exon trapping: A genetic screen to identify candidate transcribed sequences in cloned mammalian genomic DNA. Proc. Natl. Acad. Sci. U.S.A. 87: 8995 ‐ 8999.
dc.identifier.citedreference Guarente, L., Lauer, G., Roberts, T.M., and Ptashne, M. 1980. Improved methods for maximizing expression of a cloned gene: A bacterium that synthesizes rabbit β‐globin. Cell 20: 543 ‐ 553.
dc.identifier.citedreference Hanzlik, A.J., Hauser, M.A., Osemlak‐Hanzlik, M.M., and Kurnit, D.M. 1993. The recombination‐based assay demonstrates that the fragile X sequence is transcribed widely during development. Nature Genet. 3: 44 ‐ 48.
dc.identifier.citedreference Hochgeschwender, U. 1994. Identifying transcribed sequences in arrayed bacteriophage or cosmid libraries. In Current Protocols in Human Genetics ( Dracopoli, N., Haines, J.L., Korf, B., Moir, D.T., Morton, C.M., Seidman, C.E., Seidman, J.G., and Smith, D.R., eds.) pp. 6.2.1 ‐ 6.2.15. John Wiley & Sons, New York.
dc.identifier.citedreference Hochgeschwender, U., Sutcliffe, J.G., and Brennan, M.D. 1989. Construction and screening of a genomic library specific for mouse chromosome 16. Proc. Natl. Acad. Sci. U.S.A. 86: 8482 ‐ 8486.
dc.identifier.citedreference Huynh, T., Young, R.A., and Davis, R.W. 1985. Constructing and screening cDNA libraries in λgt10 and λgt11. In DNA cloning, Vol. II ( D. Glover, ed.) IRL Press, Eynsham, U.K.
dc.identifier.citedreference Ikeda, H., Aoki, K., and Naito, A. 1982. Illegitimate recombination mediated in vitro by DNA gyrase of Escherichia coli: Structure of recombinant DNA molecules. Proc. Natl. Acad. Sci. U.S.A. 79: 3724 ‐ 3728.
dc.identifier.citedreference Jankowski, S., Stewart, G.D., Buraczynska, M., Galt, J., Van Keuren, M., and Kurnit, D.M. 1990. Molecular approaches to trisomy 21. Prog. Clin. Biol. Res. 360: 79 ‐ 88.
dc.identifier.citedreference Kao, F.‐T. and Yu, J.‐W. 1991. Chromsome microdissection and cloning in human genome and genetic disease analysis. Proc. Natl. Acad. Sci. U.S.A. 88: 1844 ‐ 1848.
dc.identifier.citedreference King, S.R. and Richardson, J.P. 1986. Role of homology and pathway specificity for recombination between plasmids and bacteriophage λ. Mol. Gen. Genet. 204: 141 ‐ 147.
dc.identifier.citedreference Kurachi, S., Baldori, N., and Kurnit, D.M. 1989. Sumo 15A: A lambda plasmid that permits easy selection for and against cloned inserts. Gene 85: 35 ‐ 43.
dc.identifier.citedreference Kurnit, D.M. and Seed, B. 1990. Improved genetic selection for screening bacteriophage libraries by homologous recombination in vivo. Proc. Natl. Acad. Sci. U.S.A. 87: 3166 ‐ 3169.
dc.identifier.citedreference Lawn, R.M., Fritsch, E.H., Parker, R.C., Blake, G., and Maniatis, T. 1978. The isolation and characterization of linked δ‐ and β‐globin genes from a cloned library of human DNA. Cell 15: 1157 ‐ 1174.
dc.identifier.citedreference Liu, P., Legerski, R., and Siciliano, M.J. 1989. Isolation of human transcribed sequences from human‐rodent somatic cell hybrids. Science 246: 813 ‐ 815.
dc.identifier.citedreference Lovett, M. 1994. Direct selection of cDNAs using genomic contigs. In Current Protocols in Human Genetics ( N. Dracopoli, J.L. Haines, B. Korf, D.T. Moir, C.M. Morton, C.E. Seidman, J.G. Seidman, and D.R. Smith, eds.) pp. 6.3.1 ‐ 6.3.15. John Wiley & Sons, New York.
dc.identifier.citedreference Lovett, M., Kere, J., and Hinton, L.M. 1991. Direct selection: A method for the isolation of cDNAs encoded by large genomic regions. Proc. Natl. Acad. Sci. U.S.A. 88: 9628 ‐ 9632.
dc.identifier.citedreference Lutz, C.T., Hollifield, W.C., Seed, B., Davie, J.M., and Huang, H.V. 1987. Syrinx 2A: An improved λ phage vector designed for screening DNA libraries by recombination in vivo. Proc. Natl. Acad. Sci. U.S.A. 84: 4379 ‐ 4383.
dc.identifier.citedreference Nisson, P.E. and Watkins, P.C. 1994. Isolation of exons from cloned DNA by exon trapping. In Current Protocols in Human Genetics ( Dracopoli, N., Haines, J.L., Korf, B., Moir, D.T., Morton, C.M., Seidman, C.E., Seidman, J.G., and Smith, D.R., eds.) pp. 6.1.1 ‐ 6.1.14. John Wiley & Sons, New York.
dc.identifier.citedreference Parimoo, S., Patanjali, S.R., Shukla, H., Chaplin, D.D., and Weissman, S.M. 1991. cDNA selection: Efficient PCR approach for the selection of cDNAs encoded in large chromosomal DNA fragments. Proc. Natl. Acad. Sci. U.S.A. 88: 9623 ‐ 9627.
dc.identifier.citedreference Poustka, A., Rackwitz, H.‐R., Frischauf, A., Hohn, B., and Lehrach, H. 1984. Selective isolation of cosmid clones by homologous recombination in Escherichia coli. Proc. Natl. Acad. Sci. U.S.A. 81: 4129 ‐ 4133.
dc.owningcollname Interdisciplinary and Peer-Reviewed
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