ジゴキシゲニン標識オリゴヌクレオチドプローブ(CAC)5を用いたDNAフィンガープリント法によるネコ条虫の遺伝的変異の解析

ネコ条虫よりDNAを抽出し, ジゴキシゲニン標識オリゴヌクレオチドプローブ(CAC)5を用いたサザンハイブリダイゼーションを行ったところ, 明瞭なDNAフィンガープリント像が得られた. このDNAフィンガープリント像は, 宿主のものとは全く異なっており, また, 成虫と嚢虫の間でも差が見られなかったことから, ネコ条虫個体特異的と考えられた. 分離された地域, 宿主の異なるネコ条虫4分離継代株についてDNAフインガープリント法により分離株間の遺伝的変異を調べた. その結果, これまで感染性, 形態, 嚢虫の蛋白組成等では区別できなかった3分離株を含め, 4分離株すべてから分離株特異的なDNAフ...

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Published inJournal of Veterinary Medical Science Vol. 57; no. 2; pp. 267 - 272
Main Authors 岡本, 宗裕, 林, 正信, 上田, 裕美, 黒澤, 努, 奥, 祐三郎, 神谷, 正男
Format Journal Article
LanguageEnglish
Published 公益社団法人 日本獣医学会 1995
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ISSN0916-7250
1347-7439
DOI10.1292/jvms.57.267

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Abstract ネコ条虫よりDNAを抽出し, ジゴキシゲニン標識オリゴヌクレオチドプローブ(CAC)5を用いたサザンハイブリダイゼーションを行ったところ, 明瞭なDNAフィンガープリント像が得られた. このDNAフィンガープリント像は, 宿主のものとは全く異なっており, また, 成虫と嚢虫の間でも差が見られなかったことから, ネコ条虫個体特異的と考えられた. 分離された地域, 宿主の異なるネコ条虫4分離継代株についてDNAフインガープリント法により分離株間の遺伝的変異を調べた. その結果, これまで感染性, 形態, 嚢虫の蛋白組成等では区別できなかった3分離株を含め, 4分離株すべてから分離株特異的なDNAフインガープリント像が得られ, 互いに明瞭に区別することができた. (CAC)5は, 条虫の遺伝的変異を調べる際に非常に有用なプローブであると考えられた. 32P標識とジゴキシゲニン標識のプローブを使用した場合のフィンガープリント像を比較すると, ジゴキシゲニン標識では, バンドの強度が平均化する傾向が見られ,32P標識の場合に較べより多くの明瞭なバンドを検出することができた. 条虫の遺伝的変異を検出するにはジゴキシゲニン標識プローブがより有効と考えられた.
AbstractList ネコ条虫よりDNAを抽出し, ジゴキシゲニン標識オリゴヌクレオチドプローブ(CAC)5を用いたサザンハイブリダイゼーションを行ったところ, 明瞭なDNAフィンガープリント像が得られた. このDNAフィンガープリント像は, 宿主のものとは全く異なっており, また, 成虫と嚢虫の間でも差が見られなかったことから, ネコ条虫個体特異的と考えられた. 分離された地域, 宿主の異なるネコ条虫4分離継代株についてDNAフインガープリント法により分離株間の遺伝的変異を調べた. その結果, これまで感染性, 形態, 嚢虫の蛋白組成等では区別できなかった3分離株を含め, 4分離株すべてから分離株特異的なDNAフインガープリント像が得られ, 互いに明瞭に区別することができた. (CAC)5は, 条虫の遺伝的変異を調べる際に非常に有用なプローブであると考えられた. 32P標識とジゴキシゲニン標識のプローブを使用した場合のフィンガープリント像を比較すると, ジゴキシゲニン標識では, バンドの強度が平均化する傾向が見られ,32P標識の場合に較べより多くの明瞭なバンドを検出することができた. 条虫の遺伝的変異を検出するにはジゴキシゲニン標識プローブがより有効と考えられた.
Author 奥, 祐三郎
岡本, 宗裕
上田, 裕美
黒澤, 努
林, 正信
神谷, 正男
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9. Iwamura, Y., Irie, Y., Kominami, R., Nara, T., and Yasuraoka, K. 1991. Existence of host-related DNA sequences in the schistosome genome. Parasitology 102: 397-403.
19. Thomspon, R. C. A. 1988. Intraspecific variation and epidemiology. pp. 391-411. In: Parasitology in Focus (Melhorn, H. ed.), Springer, Berlin.
8. Iwamura, Y. and Irie, Y. 1992. Heterogeneity of hostrelated DNA sequences in schistosomes. Parasitol. Today 8: 90.
10. Jeffreys, A. J., Brookfield, J. F. Y., and Semeonoff, R. 1985. Positive identification of an immigration test-case using DNA fingerprints. Nature (Lond.) 317: 818-819.
16. Pena, S. D., Macedo, A. M., Gontijo, N. F., Medeiros, A. M., and Ribeiro, J. C. 1991. DNA bioprints: Simple nonisotopic DNA fingerprints with biotinylated probes. Electrophoresis 12: 146-152.
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3. Bierwerth, S., Kahl, G., Weigand, F., and Weising, K. 1992. Oligonucleotide fingerprinting of plant and fungal genomes: A comparison of radioactive, colorigenic and chemiluminescent detection methods. Electrophoresis 13: 115-122.
7. Iwaki, T., Nonaka, N., Okamoto, M., Oku, Y., and Kamiya, M. 1994. Developmental and morphological characteristics of Taenia taeniaeformis (Batsch, 1786) in Clethrionomys rufocanus bedfordiae and Rattus norvegicus from different geographical locations. J. Parasitol. 80: 461-467.
11. Jeffreys, A. J., Wilson, V., and Theirs, S. L. 1985. Hypervariable `minisatellite' regions in human DNA. Nature (Lond.) 314: 67-73.
2. Azuma, H., Okamoto, M., Oku, Y., and Kamiya, M. 1995. Intraspecific variation in Taenia taeniaeformis by various criteria. Parasitol. Res. 81: 103-108.
1. Ali, S., Miiller, C. R., and Epplen, J. T. 1986. DNA finger printing by oligonucleotide probes specific for simple repeats. Hum. Genet. 74: 237-243.
20. Thompson, R. C. A. and Lymbery, A. J. 1988. The nature, extent and significance of variation within the genus Echinococcus. Adv. Parasitol. 27: 209-258.
6. Gill, P., Jeffreys, A. J., and Werrett, D. J. 1985. Forensic application of DNA 'fingerprints'. Nature (Lond.) 318: 577-579.
23. Zischlr, H., Nanda, I., Schäfer, R., Schmid, M., and Epplen, J. T. 1989. Digoxigenated oligonucleotide probes specific for simiple repeats in DNA fingerprinting and hybridization in situ. Hum. Genet. 82: 227-233.
17. Rollinson, D., Walker, T. K., and Simpson, A. J. G. 1986. The application of recombinant DNA technology to problems of helminth identification. Parasitology 91: S53-S71.
12. Nara, T., Iwamura, Y., Tanaka, M., Irie, Y., and Yasuraoka, K. 1990. Dynamic changes of DNA sequences in Schistosoma mansoni in the course of development. Parasitology 100: 241-245.
15. Okamoto, M., Ito, A., Kurosawa, T., Oku, Y., Kamiya, M., and Agatsurna, T. 1995. Intraspecific variation of isoenzymes in Taenia taeniaeformis. Int. J. Parasitol. 25: 221-228.
22. Zischler, H., Hinkkanen, A., and Studer, R. 1991. Oligonucleotide fingerprinting with (CAC)5: Nonradioactive ingel hybridization and isolation of individual hypervariable loci. Electrophoresis 12: 141-146.
References_xml – reference: 15. Okamoto, M., Ito, A., Kurosawa, T., Oku, Y., Kamiya, M., and Agatsurna, T. 1995. Intraspecific variation of isoenzymes in Taenia taeniaeformis. Int. J. Parasitol. 25: 221-228.
– reference: 19. Thomspon, R. C. A. 1988. Intraspecific variation and epidemiology. pp. 391-411. In: Parasitology in Focus (Melhorn, H. ed.), Springer, Berlin.
– reference: 9. Iwamura, Y., Irie, Y., Kominami, R., Nara, T., and Yasuraoka, K. 1991. Existence of host-related DNA sequences in the schistosome genome. Parasitology 102: 397-403.
– reference: 14. Nürnberg, P., Roewer, L., Neitzel, H., Sperling, K. Pöpperl, A., Hundrieser, J., Pöche, H., Epplen, C., Zischler, H., and Epplen, J. T. 1989. DNA fingerprinting with the oligonucleotide probe (CAC)5/(GTG)5: somatic stability and germline mutations. Hum. Genet. 84: 75-78.
– reference: 1. Ali, S., Miiller, C. R., and Epplen, J. T. 1986. DNA finger printing by oligonucleotide probes specific for simple repeats. Hum. Genet. 74: 237-243.
– reference: 8. Iwamura, Y. and Irie, Y. 1992. Heterogeneity of hostrelated DNA sequences in schistosomes. Parasitol. Today 8: 90.
– reference: 20. Thompson, R. C. A. and Lymbery, A. J. 1988. The nature, extent and significance of variation within the genus Echinococcus. Adv. Parasitol. 27: 209-258.
– reference: 18. Simpson, A. J. G. and Pena, S. D. J. 1992. Host-related DNA sequences in the schistosome genome. Parasitol. Today 7: 266.
– reference: 23. Zischlr, H., Nanda, I., Schäfer, R., Schmid, M., and Epplen, J. T. 1989. Digoxigenated oligonucleotide probes specific for simiple repeats in DNA fingerprinting and hybridization in situ. Hum. Genet. 82: 227-233.
– reference: 4. Bruford, M. W., Hanotte, 0., Brookfield, J. F. Y., and Burke, T. 1992. Single-locus and multilocus DNA fingerprinting. pp. 225-269. In: Molecular Genetic Analysis of Populations (Hoelzel, A. R. ed.), Oxford University Press, New York.
– reference: 13. Nonaka, N., Iwaki, T., Okamoto, M., Ooi, H.-K., Oku, Y., Ohbayashi, M., and Kamiya, M. 1994. Infectivities of four isolates of Taenia taeniaeformis to various rodents. J. Vet. Med. Sci. 56: 565-567.
– reference: 17. Rollinson, D., Walker, T. K., and Simpson, A. J. G. 1986. The application of recombinant DNA technology to problems of helminth identification. Parasitology 91: S53-S71.
– reference: 6. Gill, P., Jeffreys, A. J., and Werrett, D. J. 1985. Forensic application of DNA 'fingerprints'. Nature (Lond.) 318: 577-579.
– reference: 22. Zischler, H., Hinkkanen, A., and Studer, R. 1991. Oligonucleotide fingerprinting with (CAC)5: Nonradioactive ingel hybridization and isolation of individual hypervariable loci. Electrophoresis 12: 141-146.
– reference: 5. Buitkamp, J., Ammer, H., and Geldermann, H. 1991. DNA fingerprinting in domestic animals. Electrophoresis 12: 169-174.
– reference: 12. Nara, T., Iwamura, Y., Tanaka, M., Irie, Y., and Yasuraoka, K. 1990. Dynamic changes of DNA sequences in Schistosoma mansoni in the course of development. Parasitology 100: 241-245.
– reference: 3. Bierwerth, S., Kahl, G., Weigand, F., and Weising, K. 1992. Oligonucleotide fingerprinting of plant and fungal genomes: A comparison of radioactive, colorigenic and chemiluminescent detection methods. Electrophoresis 13: 115-122.
– reference: 2. Azuma, H., Okamoto, M., Oku, Y., and Kamiya, M. 1995. Intraspecific variation in Taenia taeniaeformis by various criteria. Parasitol. Res. 81: 103-108.
– reference: 7. Iwaki, T., Nonaka, N., Okamoto, M., Oku, Y., and Kamiya, M. 1994. Developmental and morphological characteristics of Taenia taeniaeformis (Batsch, 1786) in Clethrionomys rufocanus bedfordiae and Rattus norvegicus from different geographical locations. J. Parasitol. 80: 461-467.
– reference: 10. Jeffreys, A. J., Brookfield, J. F. Y., and Semeonoff, R. 1985. Positive identification of an immigration test-case using DNA fingerprints. Nature (Lond.) 317: 818-819.
– reference: 16. Pena, S. D., Macedo, A. M., Gontijo, N. F., Medeiros, A. M., and Ribeiro, J. C. 1991. DNA bioprints: Simple nonisotopic DNA fingerprints with biotinylated probes. Electrophoresis 12: 146-152.
– reference: 11. Jeffreys, A. J., Wilson, V., and Theirs, S. L. 1985. Hypervariable `minisatellite' regions in human DNA. Nature (Lond.) 314: 67-73.
– reference: 21. Wolfes, R., Máthé, J., and Seitz, A. 1991. Forensic of birds of prey by DNA fingerprinting with 32P-labeled oligonucleotide probes. Electrophoresis 12: 175-180.
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Title ジゴキシゲニン標識オリゴヌクレオチドプローブ(CAC)5を用いたDNAフィンガープリント法によるネコ条虫の遺伝的変異の解析
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