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Evaluation and characterization of a porcine small intestine cDNA library: Analysis of 839 clones

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Abstract

A porcine small intestine directionally cloned cDNA library was constructed in the vector lambda Zap II. Clones were hybridized with total labeled cDNA such that putative high-copy number transcripts could be differentiated from middle- and low-copy number transcripts prior to selection and characterization by DNA sequencing. More than 2000 non-hybridizing and 242 hybridizing clones were collected. In total, 839 clones were sequenced from the 3′ end of the cDNA, and after inter-clone comparison, the unique clones were sequenced from the 5′ end of the cDNA. The 5′ data were used to query the sequence in databases and resulted in the identification of 630 different gene transcripts, of which 604 are new porcine genes. The identity of 361 transcripts could be identified from sequence comparison studies. The validity of this semi-random selection approach was verified by the identification of a large number of unique transcripts.

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References

  • Actams, M.D., Kelley, J.M., Gocayne, J.D., Dubnick, M., Polymeropoulos, M.H., Xiao, H., Merril, C.R., Wu, A., Olde, B., Moreno, R.F., Kerlavage, A.R., McCombie, W.R., Venter, J.C. (1991). Complementary DNA sequencing: expressed sequence tags and human genome project. Science 252, 1651–1656.

    Article  Google Scholar 

  • Actams, M.D., Soares, M.B., Kerlavage, A.R., Fields, C, Venter, J.C. (1993). Rapid cDNA sequencing (expressed sequence tags) from a human brain cDNA library. Nature Genet. 4, 373–380.

    Article  Google Scholar 

  • Actams, M.D., Kerlavage, A.R., Fleischmann, R.D., Fuldner, R.A., Bult, C.J. et al. (1995). Initial assessment of human gene diversity and expression patterns based upon 83 million nucleotides of cDNA sequence. Nature 377, Suppl., 3–174.

    Google Scholar 

  • Andersson, L., Haley, C.S., Ellegren, H., Knott, S.A., Johansson, M., Andersson, K., Andersson-Eklund, L., Edfors-Lilja, I., Fredholm, M., Hansson, I., Håkansson, J., Lundstrsöm, K. (1994). Genetic mapping of quantitative trait loci for growth and fatness in pigs. Science 263, 1771–1774.

    Article  PubMed  CAS  Google Scholar 

  • Archibald, A.L., Brown, F.J., Couperwhite, S., McQueen, H.A., Nicholson, D., Haley, C.S., Coppierters, W., van de Weghe, A., Stratil, A., Winterø, A.K., Fredholm, M., Larsen, N.J., Nielsen, V.H., Milan, D., Woloszyn, A.R., Dalens, M, Riquet, J., Gellin, J., Caritez, J-C, Hue, D., Burgaud, G., Ollivier, L., Bidanel, J-P., Vaiman, M., Renard, C, Geldermann, H., Davoli, R., Reyter, D., Verstege, E.J.M., Groenen, M.A.M., Davies, W., Høyheim, B., Keiserud, A., Andersson, L., Ellegren, H., Johansson, M., Marklund, L., Miller, R.J., Anderson Dear, D.V., Signer, E., Jeffreys, A.J., Moran, C, Le Tissier, P., Rothschild, M.F., Tuggle, C.K., Vaske, D., Helm, J., Liu, H.-C., Rahma, A., Yu, T.-P., Larson, R.G., Schmitz, C.B. (1995). The PiGMaP consortium linkage map of the pig (Sus scrofa). Mamm. Genome 6, 157–175.

    Article  PubMed  CAS  Google Scholar 

  • Ballabio, A. (1993). The rise and fall of positional cloning? Nature Genet. 3, 277–279.

    Article  PubMed  CAS  Google Scholar 

  • Beier, D.R. (1993). Single-strand conformation polymorphism (SSCP) analysis as a tool for genetic mapping. Mamm. Genome 4, 627–631.

    Article  PubMed  CAS  Google Scholar 

  • Bishop, J.O., Morton, J.G., Rosbash, M., Richardson, M. (1974). Three abundance classes in HeLa cell messenger RNA. Nature 250, 199–204.

    Article  PubMed  CAS  Google Scholar 

  • Colins, F.S. (1995). Positional cloning moves from perditional to traditional. Nature Genet. 9, 347–350.

    Article  Google Scholar 

  • Ellegren, H., Chowdhary, B.P., Johansson, M., Marklund, L., Fredholm, M., Gustavsson, I., Andersson, L. (1994). A primary linkage map of the porcine genome reveals a low rate of genetic recombination. Genetics 137, 1089–1100.

    PubMed  CAS  Google Scholar 

  • Firgerio, J.-M., Berthézsène, P., Garrido, P., Ortiz, E., Barthellemy, S., Vasseur, S., Sastre, B., Seleznieff, I., Dagorn, J.-C, Iovanna, J.L. (1995). Analysis of 2166 clones from a human colorectal cancer cDNA library by partial sequencing. Hum. Mol. Genet. 4, 37–43.

    Article  Google Scholar 

  • Fujii, J., Otsu, K., Zorzato, F., de Leon, S., Khanna, V.K., Weiler, J.E., O’;brien, P.J., MacLennan, D.H. (1991). Identification of a mutation in porcine ryanodine receptor associated with malignant hyperthermia. Science 253, 448–451.

    Article  PubMed  CAS  Google Scholar 

  • Gish, W., States, D.J. (1993). Identification of protein coding regions by database similarity search. Nature Genet. 3, 266–272.

    Article  PubMed  CAS  Google Scholar 

  • Höög, C. (1991). Isolation of a large number of novel mammalian genes by a differential cDNA library screening strategy. Nucleis Acids Res. 19, 6123–6127.

    Article  Google Scholar 

  • Johansson, M., Ellegren, H., Andersson, L. (1995). Comparative mapping reveals extensive linkage conservation-but with gene order rearrangements-between the pig and the human genome. Genomics 25, 682–690.

    Article  PubMed  CAS  Google Scholar 

  • Khan, A.S., Wilcos, A.S., Polymeropoulos, M.H., Hopkins, J.A., Stevens, T.J., Robinson, M., Orpana, A.K., Sikela, J.M. (1992). Single pass sequencing and physical and genetic mapping of human brain cDNAs. Nature Genet. 2, 180–185.

    Article  PubMed  CAS  Google Scholar 

  • O’;Brien, S.J., Peters, J. Searle, A.G., Womack, J.E., Johnson, P.A. and Graves, J.A.M. (1994). Report of the committee on comparative gene mapping, 1993. In Human Gene Mapping, A.J. Cuticchia, P.L. Pearson, eds. (Baltimore: The John Hopkins University Press), pp. 846–892.

    Google Scholar 

  • Pearson, W.R., Lipman, D.J. (1988). Improved tools for biological sequence comparison. Proc. Natl. Acad. sci. USA 85, 2444–2448.

    Article  PubMed  CAS  Google Scholar 

  • Rohrer, G.A., Alexander, L.J., Keele, J.W., Smith, T.P., Beattie, C.W. (1994). A microsatellite linkage map of the porcine genome. Genetics 136, 231–245.

    PubMed  CAS  Google Scholar 

  • Sambrook, J., Fritsch, E.F., Maniatis, T. (1989). Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor, N.Y.: Cold Spring Harbor Laboratory Press).

    Google Scholar 

  • Sudo, K., Chinen, K., Nakamura, Y. (1994). 2058 expressed sequence tags (ESTs) from a human fetal lung cDNA library. Genomics 24, 276–279.

    Article  PubMed  CAS  Google Scholar 

  • Takeda, J., Yano, H., Eng, S., Zeng, Y., Bell, G.I. (1993). A molecular inventory of human pancreatic islets: sequence analysis of 1000 cDNA clones. Hum. Mol. Genet. 2, 1793–1798.

    Article  PubMed  CAS  Google Scholar 

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Winterø, A.K., Fredholm, M. & Davies, W. Evaluation and characterization of a porcine small intestine cDNA library: Analysis of 839 clones. Mammalian Genome 7, 509–517 (1996). https://doi.org/10.1007/s003359900153

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  • DOI: https://doi.org/10.1007/s003359900153

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