Araştırma Makalesi
BibTex RIS Kaynak Göster
Yıl 2019, Cilt: 4 Sayı: 2, 134 - 138, 31.08.2019
https://doi.org/10.35229/jaes.550588

Öz

Kaynakça

  • Bailes, S. M., Devers, J. J., Kirby, J. D. & Rhoads, D. D. (2007). An inexpensive, simple protocol for dna isolation from blood for high-throughput genotyping by polymerase chain reaction or restriction endonuclease digestion. Poultry Science, 86(1), 102–106. doi: 10.1093/ps/86.1.102.
  • Barwise, A. H. & Walker, I. O. (1970). Studies on the DNA of a virüs from Galleria mellonella. Febs Letters, 6(1), 13-16. doi: 10.1016/0014-5793(70)80028-X.
  • Bio-rad laboratories (2018). Insta Gene Matrix: Prepare DNA templates for PCR with no phenol/chloroform extractions and no deproteinization steps. Tech note 1810. Erişim tarihi: 05.04.2018.
  • Bozkaya, F. (2012). DNA izolasyonunda Fenol-Kloroform yerine Potasyum-Asetat Kullanımının DNA miktarı ve Kalitesine Etkisi”, Harran Üniversitesi Veteriner Fakültesi Dergisi, 1(2), 92-96.
  • Bronskill, J. K. (1961). A cage to simpllify the rearing of greater wax moth, Galleria mellonella (Pyralidae). Journal of Lepidopterist’s Society, 15(2), 102-104.
  • Chen, H. M., Wang, W., Smith, D. & Chan, S.C. (1997). Effects of the antibacterial peptide cecropin B and its analogs, cecropins B-1 and B-2, on liposomes, bacteria and cancer cells. Biochimica et Biophysica Acta, 1336(2), 171-179. doi: 10.1016/S0304-4165(97)00024-X.
  • Cherepenko, E. I., Galkin, A. P., Lebedev, V. R. & Kok, I. P. (1982). A non-damaging procedure for DNA isolation from insect cell nuclei. Ukrainian Biochemical Journal, 54(3), 316-321.
  • Cytrynska, M., Mak, P., Zdybicka-Barabas, A., Suder, P. & Jakubowicz, T. (2007). Purification and characterization of eight peptides from Galleria mellonella immune hemolymph. Peptides, 28(3), 533-546. doi: 10.1016/j.peptides.2006.11.010.
  • Cytrynska, M., Zdybicka-Barabas, A., Jablonski, P. & Jakubowicz, T. (2001). Detection of antibacterial polypeptide activity in situ after sodium dodecyl sulfate-polyacrylamide gel elecrophoresis. Analytical Biochemistry, 299(2), 274-276. doi: 10.1006/abio.2001.5422.
  • Emre, I., Kayis, T., Coskun, M., Dursun, O. & Cogun, H. Y. (2013). Changes in antioxidative enzyme activity, glycogen, lipid, protein, and malondialdehyde content in cadmium-treated Galleria mellonella larvae. Annals of the Entomological Society of America, 106(3), 371–377. doi: 10.1603/AN12137.
  • Kayis, T., Coskun, M., Dursun, O. & Emre, I. (2015). Alterations in antioxidant enzyme activity, lipid peroxidation and ion balance induced by Dichlorvos in Galleria mellonella L. Annals of the Entomological Society of America, 108(4), 570–574. doi: 10.1093/aesa/sav038.
  • Kim, C. H., Lee, J. H., Kim, I., Seo, S. J., Son, S. M. & Lee, K. Y. (2004). Purification and cDNA cloning of a cecropin-like peptide from the great wax moth, Galleria mellonella. Molecules and Cells, 17(2), 262-266.
  • Lahiri, D. K., Bye, S., Nurnberger, J. I., Hodes, M. E. & Crisp, M. (1992). A non-organic and non-enzymatic extraction method gives higher yields of genomic DNA from whole-blood samples than do nine other methods tested. Journal of Biochemical and Biophysical Methods, 25(4), 193–205. doi: 10.1016/0165-022X(92)90014-2.
  • Laird, P. W., Zijderveld, A., Linders, K., Rudnicki, M. A., Jaenisch, R. & Berns, A. (1991). Simplified mammalian DNA isolation procedure. Nucleic Acids Research, 19(15), 4293.
  • Lamberty, M., Ades, S., Uttenweiler-Joseph, S., Brookhart, G., Bushey, D., Hoffmann, J. A. & Bulet, P. (1999). Insect immunity. Isolation from the lepidopteran Heliothis virescens of a novel insect defensin with potent antifungal activity. Journal of Biological Chemistry, 274(14), 9320-9326. doi: 10.1074/jbc.274.14.9320.
  • Mak, P., Zdybicka-Barabas, M. & Cytynska, A. (2010). A different repertoire of Galleria mellonella antimicrobial peptides in larvae challenged with bacteria and fungi. Developmental & Comparative Immunology, 34(10), 1129-36. doi: 10.1016/j.dci.2010.06.005.
  • Mukherjee, K., Altincicek, B., Hain, T., Domann, E., Vilcinskas, A. & Chakraborty, T. (2010). Galleria mellonella as a model system for studying Listeria pathogenesis. Applied and Environmental Microbiololgy, 76(1), 310-317. doi: 10.1128/AEM.01301-09.
  • Nale, Y. J., Chutia, M., Carr, P., Hickenbotham, P. T. & Clokie, M .R. J. (2016). Get in early: Biofilm and wax moth (Galleria mellonella) models reveal new insights into the therapeutic potential of Clostridium difficile bacteriophages. Frontiers in Microbiology, 7:1383. doi: 10.3389/fmicb.2016.01383.
  • Ponnuvel, K. M. & Yamakawa, M. (2002). Immune responses against bacterial infection in Bombyx mori and regulation of host gene expression. Current Science, 83(4), 447-454.
  • Rowley, A. F. & Ratcliffe, N. A. (1981). Insects, In: Invertebrate Blood Cells, ed. Ratcliffe N.A., Rowley A.F., Academic Press, London, 2, 421-488.
  • Sambrook, J. & Russell, D. W. (2001). Commonly Used Techniques in Molecular Cloning, Appendix 8 in Molecular Cloning, 3rd edition. New York: Cold Spring Harbor, 2208 p.
  • Sambrook, J., Fritsch, E. F. & Maniatis, T. (1989). Molecular Cloning: A laboratory manual, Cold Spring Harbor Laboratory Press. New York.
  • Strand, M. R. (2008). The insect cellular immune response. Insect Science, 15, 1-14. doi: 10.1111/j.1744-7917.2008.00183.x.
  • Strøm, G. E. A., Tellevik, M. G., Hanevik, K., Langeland, N. & Blomberg, B. (2014). Comparison of four methods for extracting DNA from dried blood on filter paper for PCR targeting the mitochondrial Plasmodium genome. Transactions of The Royal Society of Tropical Medicine and Hygiene, 108(8):488-94. doi: 10.1093/trstmh/tru084.
  • Wagley, S., Borne, R., Harrison, S., Baker-Austin, C., Ottaviani, D., Leoni, F., Vuddhakul, V. & Titball, R. W. (2018). Galleria mellonella as an infection model to investigate virulence of Vibrio parahaemolyticus. Vilulence, 9(1), 197-207. doi: 10.1080/21505594.2017.1384895.
  • Zasloff, M. (2002). Antimicrobial peptides of multicellular organisms. Nature, 415(6870), 389-395. doi: 10.1038/415389a.

Model Organizma Galleria mellonella’ da Hemolenf ve Dokudan Düşük Maliyetli ve Basit DNA İzolasyon Metodu

Yıl 2019, Cilt: 4 Sayı: 2, 134 - 138, 31.08.2019
https://doi.org/10.35229/jaes.550588

Öz

Bu
çalışmanın amacı, rutin laboratuvar çalışmalarında sıklıkla kullanılan ekonomik
kimyasalları ve sarf malzemeleri kullanarak dokudan ve vücut sıvılarından mevcut
izolasyon yöntemlerine alternatif, polimeraz zincir reaksiyonu (PZR)
işlemlerinde kullanılmak üzere gDNA izolasyon yöntemi oluşturmaktır. Bu amaçla
model organizma olarak Galleria
mellonella
L. (Lepidoptera: Pyralidae) kullanılmıştır.
Çalışmada,
oluşturduğumuz yöntemle karşılaştırma yapabilmek için iki farklı DNA izolasyon
yöntemi de uygulanmıştır. İzolasyonu yapılan DNA’ ların miktar ve saflığı 260 nm
ve 260 nm/280 nm’ de spektrofotometrik ölçümlerle belirlenmiş ve agaroz jel
elektroforezi ile kontrolleri yapılmıştır. Fenolsüz ve santrifüjsüz yöntem,
InstaGene matrix yöntemi ve bazı modifikasyonlar yaparak oluşturduğumuz DNA
izolasyon yöntemi ile hemolenf örneğinden elde edilen A260 değerleri
sırasıyla 0,032, 0,157, 0,432; doku örneğinden elde edilen A260
değerleri ise sırasıyla 0,291, 0,121, 0,346 bulunmuştur. Saflık değerleri ise
hemolenf grubunda sırasıyla 1,030, 1,401, 1,878; doku grubunda ise 1,785,
1,690, 2,011 olarak tespit edilmiştir.



Sonuç olarak modifiye edilerek oluşturduğumuz
yöntem ile hem vücut sıvısından hem de dokudan yeterli miktar ve kalitede DNA
elde edilebildiği tespit edilmiştir.

Kaynakça

  • Bailes, S. M., Devers, J. J., Kirby, J. D. & Rhoads, D. D. (2007). An inexpensive, simple protocol for dna isolation from blood for high-throughput genotyping by polymerase chain reaction or restriction endonuclease digestion. Poultry Science, 86(1), 102–106. doi: 10.1093/ps/86.1.102.
  • Barwise, A. H. & Walker, I. O. (1970). Studies on the DNA of a virüs from Galleria mellonella. Febs Letters, 6(1), 13-16. doi: 10.1016/0014-5793(70)80028-X.
  • Bio-rad laboratories (2018). Insta Gene Matrix: Prepare DNA templates for PCR with no phenol/chloroform extractions and no deproteinization steps. Tech note 1810. Erişim tarihi: 05.04.2018.
  • Bozkaya, F. (2012). DNA izolasyonunda Fenol-Kloroform yerine Potasyum-Asetat Kullanımının DNA miktarı ve Kalitesine Etkisi”, Harran Üniversitesi Veteriner Fakültesi Dergisi, 1(2), 92-96.
  • Bronskill, J. K. (1961). A cage to simpllify the rearing of greater wax moth, Galleria mellonella (Pyralidae). Journal of Lepidopterist’s Society, 15(2), 102-104.
  • Chen, H. M., Wang, W., Smith, D. & Chan, S.C. (1997). Effects of the antibacterial peptide cecropin B and its analogs, cecropins B-1 and B-2, on liposomes, bacteria and cancer cells. Biochimica et Biophysica Acta, 1336(2), 171-179. doi: 10.1016/S0304-4165(97)00024-X.
  • Cherepenko, E. I., Galkin, A. P., Lebedev, V. R. & Kok, I. P. (1982). A non-damaging procedure for DNA isolation from insect cell nuclei. Ukrainian Biochemical Journal, 54(3), 316-321.
  • Cytrynska, M., Mak, P., Zdybicka-Barabas, A., Suder, P. & Jakubowicz, T. (2007). Purification and characterization of eight peptides from Galleria mellonella immune hemolymph. Peptides, 28(3), 533-546. doi: 10.1016/j.peptides.2006.11.010.
  • Cytrynska, M., Zdybicka-Barabas, A., Jablonski, P. & Jakubowicz, T. (2001). Detection of antibacterial polypeptide activity in situ after sodium dodecyl sulfate-polyacrylamide gel elecrophoresis. Analytical Biochemistry, 299(2), 274-276. doi: 10.1006/abio.2001.5422.
  • Emre, I., Kayis, T., Coskun, M., Dursun, O. & Cogun, H. Y. (2013). Changes in antioxidative enzyme activity, glycogen, lipid, protein, and malondialdehyde content in cadmium-treated Galleria mellonella larvae. Annals of the Entomological Society of America, 106(3), 371–377. doi: 10.1603/AN12137.
  • Kayis, T., Coskun, M., Dursun, O. & Emre, I. (2015). Alterations in antioxidant enzyme activity, lipid peroxidation and ion balance induced by Dichlorvos in Galleria mellonella L. Annals of the Entomological Society of America, 108(4), 570–574. doi: 10.1093/aesa/sav038.
  • Kim, C. H., Lee, J. H., Kim, I., Seo, S. J., Son, S. M. & Lee, K. Y. (2004). Purification and cDNA cloning of a cecropin-like peptide from the great wax moth, Galleria mellonella. Molecules and Cells, 17(2), 262-266.
  • Lahiri, D. K., Bye, S., Nurnberger, J. I., Hodes, M. E. & Crisp, M. (1992). A non-organic and non-enzymatic extraction method gives higher yields of genomic DNA from whole-blood samples than do nine other methods tested. Journal of Biochemical and Biophysical Methods, 25(4), 193–205. doi: 10.1016/0165-022X(92)90014-2.
  • Laird, P. W., Zijderveld, A., Linders, K., Rudnicki, M. A., Jaenisch, R. & Berns, A. (1991). Simplified mammalian DNA isolation procedure. Nucleic Acids Research, 19(15), 4293.
  • Lamberty, M., Ades, S., Uttenweiler-Joseph, S., Brookhart, G., Bushey, D., Hoffmann, J. A. & Bulet, P. (1999). Insect immunity. Isolation from the lepidopteran Heliothis virescens of a novel insect defensin with potent antifungal activity. Journal of Biological Chemistry, 274(14), 9320-9326. doi: 10.1074/jbc.274.14.9320.
  • Mak, P., Zdybicka-Barabas, M. & Cytynska, A. (2010). A different repertoire of Galleria mellonella antimicrobial peptides in larvae challenged with bacteria and fungi. Developmental & Comparative Immunology, 34(10), 1129-36. doi: 10.1016/j.dci.2010.06.005.
  • Mukherjee, K., Altincicek, B., Hain, T., Domann, E., Vilcinskas, A. & Chakraborty, T. (2010). Galleria mellonella as a model system for studying Listeria pathogenesis. Applied and Environmental Microbiololgy, 76(1), 310-317. doi: 10.1128/AEM.01301-09.
  • Nale, Y. J., Chutia, M., Carr, P., Hickenbotham, P. T. & Clokie, M .R. J. (2016). Get in early: Biofilm and wax moth (Galleria mellonella) models reveal new insights into the therapeutic potential of Clostridium difficile bacteriophages. Frontiers in Microbiology, 7:1383. doi: 10.3389/fmicb.2016.01383.
  • Ponnuvel, K. M. & Yamakawa, M. (2002). Immune responses against bacterial infection in Bombyx mori and regulation of host gene expression. Current Science, 83(4), 447-454.
  • Rowley, A. F. & Ratcliffe, N. A. (1981). Insects, In: Invertebrate Blood Cells, ed. Ratcliffe N.A., Rowley A.F., Academic Press, London, 2, 421-488.
  • Sambrook, J. & Russell, D. W. (2001). Commonly Used Techniques in Molecular Cloning, Appendix 8 in Molecular Cloning, 3rd edition. New York: Cold Spring Harbor, 2208 p.
  • Sambrook, J., Fritsch, E. F. & Maniatis, T. (1989). Molecular Cloning: A laboratory manual, Cold Spring Harbor Laboratory Press. New York.
  • Strand, M. R. (2008). The insect cellular immune response. Insect Science, 15, 1-14. doi: 10.1111/j.1744-7917.2008.00183.x.
  • Strøm, G. E. A., Tellevik, M. G., Hanevik, K., Langeland, N. & Blomberg, B. (2014). Comparison of four methods for extracting DNA from dried blood on filter paper for PCR targeting the mitochondrial Plasmodium genome. Transactions of The Royal Society of Tropical Medicine and Hygiene, 108(8):488-94. doi: 10.1093/trstmh/tru084.
  • Wagley, S., Borne, R., Harrison, S., Baker-Austin, C., Ottaviani, D., Leoni, F., Vuddhakul, V. & Titball, R. W. (2018). Galleria mellonella as an infection model to investigate virulence of Vibrio parahaemolyticus. Vilulence, 9(1), 197-207. doi: 10.1080/21505594.2017.1384895.
  • Zasloff, M. (2002). Antimicrobial peptides of multicellular organisms. Nature, 415(6870), 389-395. doi: 10.1038/415389a.
Toplam 26 adet kaynakça vardır.

Ayrıntılar

Birincil Dil Türkçe
Bölüm Makaleler
Yazarlar

Emre Gülsu 0000-0002-3800-7713

Tamer Kayış 0000-0002-1769-2133

Mustafa Coşkun 0000-0001-5044-6295

Yayımlanma Tarihi 31 Ağustos 2019
Gönderilme Tarihi 8 Nisan 2019
Kabul Tarihi 12 Haziran 2019
Yayımlandığı Sayı Yıl 2019 Cilt: 4 Sayı: 2

Kaynak Göster

APA Gülsu, E., Kayış, T., & Coşkun, M. (2019). Model Organizma Galleria mellonella’ da Hemolenf ve Dokudan Düşük Maliyetli ve Basit DNA İzolasyon Metodu. Journal of Anatolian Environmental and Animal Sciences, 4(2), 134-138. https://doi.org/10.35229/jaes.550588


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