Abstract
As more and more genes are cloned and sequenced, it is apparent that nearly all genes are related to other genes. Similar genes are grouped into families, such as the collagen and globin gene families. There are also gene superfamilies. Gene superfamilies are composed of genes that have areas of high homology and areas of high divergence. Examples of gene super families include the oncogenes, homeotic genes, and the myosin genes. In most cases, the different members of a gene family carry out related functions. A detailed protocol for the cloning by degenerate oligonucleotide polymerase chain reaction (PCR) of members of the Aquaporin family of membrane water channels (1, 2) is discussed here.
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References
Reizer, J., Reizer, A., and Saier, M. H., Jr. (1993) The MIP family of integral membrane channel proteins: sequence comparisons, evolutionary relationships, reconstructed pathways of evolution, and proposed functional differentiation of the two repeated halves of the proteins. Cut. Rev. Biochem. Mol. Biol. 28, 235–257.
Knepper, M. A. (1994) The aquaporin family of molecular water channels. Proc. Natl. Acad. Sci. USA. 91, 6255–6258.
Preston, G. M. (1993) Use of degenerate oligonucleotide primers and the polymerase chain reaction to clone gene family members, in Methods in Molecular Biology, vol. 15, PCR Protocols: Current Methods and Applications. (White, B. A., ed.), Humana, Totowa, NJ, pp 317–337.
Preston, G. M and Agre, P. (1991) Isolation of the cDNA for erythrocyte integral membrane protein of 28 kilodaltons: member of an ancient channel family. Proc. Natl. Acad. Sci. USA. 88, 11,110–11,114.
Wada, K.-N., Aota, S.-I., Tsuchiya, R., Ishibashi, F., Gojobori, T., and Ikemura, T. (1990) Codon usage tabulated from the GenBank genetic sequence data. Nucleic Acids Res. 18, 2367–2411.
Preston, G. M., Carroll, T. P., Guggino, W.B., and Agre, P. (1992) Appearance of water channels in Xenopus oocytes expressing red cell CHIP28 protein. Science. 256, 385–387.
Chrispeels, M. J. and Agre, P. (1994) Aquaporins: water channel proteins of plant and animal cells. TIBS 19, 421–425.
Jung, J. S., Bhat, B. V., Preston, G. M., Guggino, W. B., Baraban, J. M., and Agre P. (1994) Molecular characterization of an aquaporin cDNA from brain. candidate osmoreceptor and regulator of water balance. Proc. Natl. Acad. Sci. USA. 91, 13,052–13,056.
Raina, S., Preston, G. M., Guggino, W. B., and Agre, P. (1995) Molecular cloning and characterization of an aquaporin cDNA from salivary, lacrimal and respiratory tissues. J. Biol. Chem. 270, 1908–1912.
Knoth, K., Roberds, S., Poteet, C., and Tamkun, M. (1988) Highly degenerate, inosine-containing primers specifically amplify rare cDNA using the polymerase chain reaction. Nucleic Acids Res. 16, 10,932.
Chérif-Zahar, B., Bloy, C., Kim, C. L. V., Blanchard, D., Bailly, P., Hermand, P., Salmon, C., Cartron, J.-P., and Colin, Y. (1990) Molecular cloning and protein structure of a human blood group Rh polypeptide. Proc. Natl. Acad. Sci. USA., 87, 6243–6247.
Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989) Molecular Cloning A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
Ausubel, F. M., Brent, R., Kingston, R. E., Moore, D. D., Seidman, J. G., Smith, J. A., and Struhl, K. (eds.) (1994) Current Protocols in Molecular Biology, Greene Publishing/Wiley-Interscience, New York.
Wistow, G. J., Pisano, M. M., and Chepelinsky, A. B. (1991) Tandem sequence repeats in transmembrane channel proteins. TIBS 16, 170–171.
Jung, J. S., Preston, G. M., Smith, B. L., Guggino, W. B., and Agre P. (1994) Molecular structure of the water channel through Aquaporin CHIP: the hourglass model. J. Biol. Chem. 269, 14,648–14,654.
Nielsen, S., Smith, B. L., Christensen, E. I., and Agre, P. (1993) Distribution of the aquaporm CHIP in secretory and resorptive epithelia and capillary endothelia. Proc. Natl. Acad. Sci. USA. 90, 7275–7279.
Smith, B. L., Preston, G. M., Spring, F. A., Anstee, D. J., and Agre, P. (1994) Human red cell Aquaporin CHIP, I. molecular characterization of ABH and Colton blood group antigens. J. Clin. Invest. 94, 1043–1049.
Preston, G. M., Smith, B. L., Zeidel, M. L., Moulds, J. J., and Agre P. (1994) Mutations in aquaporin-1 in phenotypically normal humans without functional CHIP water channels. Science. 265, 1585–1587.
Weyant, R. S., Edmonds, P., and Swammathan, B. (1990) Effects of ionic and nonionic detergents on the Tag polymerase. BioTechniques. 9, 308–309.
Bookstein, R., Lai, C.-C., To, H., and Lee, W.-H (1990) PCR-based detection of a polymorphic Bam HI site in intron 1 of the human retinoblastoma (RB) gene. Nucleic Acids Res. 18, 1666.
Hung, T., Mak, K., and Fong, K. (1990) A specificity enhancer for polymerase chain reaction. Nucleic Acids Res. 18, 4953.
Zhao, Z.-Y and Joho, R. H. (1990) Isolation of distantly related members in a multigene family using the polymerase chain reaction technique. Biochem. Biophys. Res. Commun. 167, 174–182.
Aelst, L. V., Hohmann, S., Zimmermann, F. K., Jans, A. W. H., and Thevelein, J. M. (1991) A yeast homologue of the bovine lens fiber MIP gene family complements the growth defect of a Saccharomyces cerevisiae mutant on fermentable sugars but not its defect in glucose-induced RAS-mediated cAMP signalling. EMBO J. 10, 2095–2104.
Muramatsu, S. and Mizuno, T. (1989) Nucleotide sequence of the region encompassing the glpKF operon and its upstream region containing a bent DNA sequence of Escherichia coli. Nucleic Acids Res. 17, 4378.
Höfte, H., Hubbard, L., Retzer, J., Ludevid, D., Herman, E. M., and Chrispeels, M. J. (1992) Vegetative and seed-specific forms of Tonoplast Intrinsic Protein in the vacuolar membrane of Arabidopsis thaliana. Plant Physiol. 99, 561–570.
Fushimi, K., Uchida, S., Hara, Y., Hirata, Y., Marumo, F., and Sasaki, S. (1993) Cloning and expression of apical membrane water channel of rat kidney collecting tubule. Nature. 361, 549–552.
Gorin, M. B., Yancey, S. B., Cline, J., Revel, J.-R., and Horwitz, J. (1984) the major intrinsic protein (MIP) of the bovine lens fiber membrane: characterization and structure based on cDNA cloning. Cell. 39, 49–59.
Guerrero, F. D., Jones, J. T., and Mullet, J. E. (1990) Turgor-responsive gene transcription and RNA levels increase rapidly when pea shoots are wilted. Sequence and expression of three induced genes. Plant Mol. Biol. 15, 11–26.
Rao, Y., Jan, L. Y., and Jan, Y. N. (1990) Similarity of the product of the Drosophila neurogenic gene big brain to transmembrane channel proteins. Nature. 345, 163–167.
Fortin, M. G., Morrison, N. A., and Verma, D. P. S. (1987) Nodulin-26, a peribacteroid membrane nodulin is expressed independently of the development of the peribacteroid compartment. Nucleic Acids Res. 15, 813–824.
Feng, D.-F. and Doolittle, R. F. (1990) Progressive alignment and phylogenetic tree construction of protein sequences, in Methods in Enzymology, vol. 183, Molecular Evolution: Computer Analysis of Protein and Nucleic Acid Sequences. (Doolittle, R. F., ed.), Academic, New York, pp. 375–387.
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© 1997 Humana Press Inc., Totowa, NJ
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Preston, G.M. (1997). Cloning Gene Family Members Using the Polymerase Chain Reaction with Degenerate Oligonucleotide Primers. In: Cowell, I.G., Austin, C.A. (eds) cDNA Library Protocols. Methods in Molecular Biology™, vol 69. Humana Press. https://doi.org/10.1385/0-89603-383-X:97
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DOI: https://doi.org/10.1385/0-89603-383-X:97
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