Research Article
BibTex RIS Cite

Kıvırcık Kuzularda PCR-RFLP Metodu Kullanılarak Calpastatin Geninin MspI ve NcoI Enzimleri ile Genetik Çeşitliliğinin Belirlenmesi

Year 2018, Volume: 44 Issue: 1, 39 - 43, 16.01.2018

Kozet Avanus

Abstract

Bu çalışmanın amacı Kıvırcık kuzularda
PCR-RFLP yöntemi ile MspI ve NcoI enzimleri kullanılarak kalpastatin geninin
genetik çeşitliliğini belirlemektir. Kırklareli ilinde bulunan sekiz farklı
çiftlikten toplanan Kıvırcık kuzularının (n=153) kan örnekleri DNA izolasyonu için
kullanılmıştır. PCR amplifikasyonundan sonra, ürünler %2’lik ve %3’lük agaroz
jel elektroforezinde M ve N allellerini ayırt etmek için MspI ve NcoI enzimleri
ile kesilmiştir. M ve N allellerinin frekansı MspI lokusunda sırasıyla %88,2 ve
%11,8, NcoI lokusunda %98,7 ve %1,3 olarak bulunmuştur. MspI lokusu için MM, MN
ve NN genotiplerinin frekansları sırasıyla 0,77, 0,22 ve 0,01 olarak tespit
edilmiştir. NcoI lokusu için MM ve MN genotiplerinin frekansları sırasıyla 0,97
ve 0,03 olarak belirlenmiştir, NN genotipi ise gözlenmemiştir. Gözlenen
heterozigotluk MspI lokusu için (0,22), NcoI lokusundan (0,03) daha yüksek
olarak bulunmuştur. Kırklareli ilinde yetiştirilen Kıvırcık kuzuları MspI ve
NcoI lokusları için Hardy-Weinberg dengesinde bulunmuştur. 

Keywords

Kalpastatin yerli koyun Kıvırcık genetik varyasyon

References

  • Aali, M., Moradi-Shahrbabak, M., Sadeghi, M., 2014. Detecting novel SNPs and breed-specific haplotypes at calpastatin gene in Iranian fat- and thin-tailed sheep breeds and their effects on protein structure. Gene 537, 132-139.
  • Asadi, N., Nanekarani, S., Khederzadeh, S., 2014. Genotypic frequency of calpastatin gene in lori sheep by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. African Journal of Biotechnology 13, 1952-1954.
  • Ata, N., Cemal, I., 2008. Calpastatin gene polymorphism in Çine Çapari and Karya sheep. Scientific Papers Series D Animal Science 56, 48-51.
  • Avanus, K., 2015. Genetic Variability of CAST Gene in Native Sheep Breeds of Turkey. Kafkas Üniversitesi Veteriner Fakültesi Dergisi 21(6), 789-794.
  • Azari, M.A., Dehnavi, E., Yousefi, S., Shahmohamadi, L., 2012. Polymorphism of calpastatin , calpain and myostatin genes in native Dalagh sheep in Iran. Slovak Journal of Animal Science 12, 1-6.
  • Byun, S.O., Zhou, H., Forrest, R.H.J., Frampton, C.M., Hickford, J.G.H., 2008. Association of the ovine calpastatin gene with birth weight and growth rate to weaning. Animal Genetics 39, 572-573.
  • Casas, E., White, S.N., Wheeler, T.L., Shackelford, S.D., Koohmaraie, R., Riley, D.G., Smith, T.P.L., 2006. Effects of calpastatin and μ-calpain markers in beef cattle on tenderness traits. Journal of Animal Science 84, 520-525.
  • Chung, H., Davis, M., 2012a. Effects of genetic variants for the calpastatin gene on calpastatin activity and meat tenderness in Hanwoo (Korean cattle). Meat Science 90; 711-714.
  • Chung, H., Davis, M., 2012b. PCR-RFLP of the ovine calpastatin gene and its association with growth. Asian Journal of Animal Science & Veterinary Advances 7, 641-652
  • Ciobanu, D.C., Bastiaansen, J.W.M., Lonergan, S.M., Thomsen, H., Dekkers, J.C.M., Rothschild, M.F., 2004. New alleles in calpastatin gene are associated with meat quality traits in pigs. Journal of Animal Science 82, 2829-2839.
  • Curi, R., Chardulo, L.L., Mason, M.C., Arrigoni, M.D.B., Silveira, C., Oliveira, H.N., 2009. Effect of single nucleotide polymorphisms of CAPN1 and CAST genes on meat traits in Nellore beef cattle (Bos indicus) and in their crosses with Bos taurus. Animal Genetics 40, 456-462.
  • Dagong, M.I., Sumantri, C., Noor, R.R., Herman, R., Yamin, M., 2011. Genetic polymorphisms of the coding region (exon 6) of calpastatin in Indonesian sheep. Media Peternakan 34, 190-195.
  • Dehnavi, E., Ahani Azari, M., Hasani, S., Nassiry, M.R., Mohajer, M., Khan, A.R., Shahmohamadi, L., Yousefi, S., 2012. Association between yearling weight and calpastatin and calpain loci polymorphism in Iranian Zel sheep. Iranian Journal of Applied & Animal Science 2, 131-135.
  • Djadid, N.D., Nikmard, M., Zakeri, S., Gholizadeh, S., 2011. Characterization of calpastatin gene in Iranian Afshari sheep. Iranian Journal of Biotechnology 9(2), 145-149.
  • Greguła-Kania, M., 2011. New allelic variant of the ovine calpastatin gene. African Journal of Biotechnology 10, 13082-13085.
  • Khan, S., Riaz, M.N., Ghaffar, A., Ullah, M.F., 2012. Calpastatin (CAST) gene polymorphism and its association with average daily weight gain in Balkhi and Kajli sheep and Beetal goat breeds. Pakistan Journal of Zoology 44(2), 377-382.
  • Khederzadeh, S., 2011. Polymorphism of calpastatin gene in crossbreed Dalagh sheep using PCR-RFLP. African Journal of Biotechnology 10, 10839-10841.
  • Mohammadi, M., Nasiri, M.T.B., Alami-Saeid, K., Fayazi, J., Mamoee, M., Sadr, A.S., 2008. Polymorphism of calpastatin gene in Arabic sheep using PCR- RFLP. African Journal of Biotechnology 7(15), 2682-2684.
  • Nanekarani, S., Khederzadeh, S., Kaftarkari, A.M., 2011. Genotypic frequency of calpastatin gene in Atabi sheep by PBR method. In: Proc. International Conference on Food Engineering and Biotechnology IPCBEE, Bangkok, Thailand, 9, 189-192.
  • Nassiry, M.R., Tahmoorespour, M., Javadmanesh, A., Soltani, M., Far, S.F., 2006. Calpastatin polymorphism and its association with daily gain in Kurdi sheep. Iranian Journal of Biotechnology 4(3), 188-192.
  • Page, B.T., Casas, E., Heaton, M.P., Cullen, N.G., Hyndman, D.L., Morris, C.A., Smith, T.P.L., 2002. Evaluation of single-nucleotide polymorphisms in CAPN1 for association with meat tenderness in cattle. Journal of Animal Science 80, 3077-3085.
  • Palmer, B.R., Roberts, N., Hickford, J.G., Bickerstaffe, R., 1998. PCR-RFLP for MspI and NcoI in the ovine calpastatin gene. Journal of Animal Science 76, 1499-1500.
  • Palmer, B.R., Su, H.Y., Roberts, N., Hickford, J.G., Bickerstaffe, R., 2000. Single nucleotide polymorphisms in an intron of the ovine calpastatin gene. Animal Biotechnology 11, 63-67.
  • Ranjbari, M., Hashemi, A., Mardani, K., Darvishzadeh, R., 2012. Allelic polymorphism of Makoei sheep calpastatin gene identified by polymerase chain reaction and single strand conformation polymorphism. Journal of Agricultural Science and Technology 14, 533-538.
  • Shahroudi, F.E., Nassiry, M.R., Valizadh, R., Heravi, A., 2006. Genetic poly-morphism at MTNR1A, CAST and CAPN loci in Iranian Karakul sheep. Iranian Journal of Biotechnology 4(2), 117-122.
  • Suleman, M., Khan, S.U., Riaz, M.N., Yousaf, M., Shah, A., Ishaq, R., 2012. Calpastatin (CAST) gene polymorphism in Kajli, Lohi and Thalli sheep breeds. African Journal of Biotechnology 4, 10655-10660.
  • Szkudlarek-Kowalczyk, M., Wiśniewska, E., Mroczkowski, S., 2011. Polymorphisms of calpastatin gene in sheep. Journal of Central European Agriculture 12, 425-432.
  • Yeh, F., Yang, R.C., Boyle, T., 2000. Popgene (v.1.32) Microsoft Windows-based freeware for Population Genetic Analysis: http://www.ualberta.ca/~fyeh/Pop32.exe (Accessed 07.09.2017)
  • Yılmaz, O., Cemal, İ., Karaca, O., Ata, N., 2014a. Association of Calpastatin (CAST) gene polymorphism with weaning weight and ultrasonic measurements of loin eye muscle in Kivircik lambs. Kafkas Universitesi Veteriner Fakültesi Dergisi 20, 675-680.
  • Yılmaz, O., Sezenler, T., Ata, N., Yaman, Y., Cemal, İ., Karaca, O., 2014b. Polymorphism of the ovine calpastatin gene in some Turkish sheep breeds. Turkish Journal of Veterinary & Animal Science 38, 354-357.
  • Zhou, H., Hickford, J.G.H., Gong, H., 2007. Polymorphism of the ovine calpastatin gene. Molecular and Cellular Probes 21, 242-244.

Determining Genetic Variation of Calpastatin Gene with MspI and NcoI Enzymes by Using PCR-RFLP Method in Kivircik Lambs

Year 2018, Volume: 44 Issue: 1, 39 - 43, 16.01.2018

Kozet Avanus

Abstract

The aim of this study was to determine the
genetic variation of calpastatin gene by using PCR-RFLP method with MspI and
NcoI enzymes in Kivircik lambs. Blood samples of Kivircik lambs (n=153) that
were collected from eight different farms located in Kirklareli province, were
used for DNA isolation. After PCR amplification, products were digested with
MspI and NcoI enzymes to differentiate M and N alleles on 2% and 3% agarose gel
electrophoresis. The frequency of M and N alleles were found 88.2% and 11.8%
for MspI locus and 98.7% and 1.3% for NcoI locus respectively. The frequencies
of MM, MN and NN genotypes were identified 0.77, 0.22 and 0.01 respectively for
MspI locus. The frequencies of MM and MN genotypes were determined 0.97 and
0.03 respectively for NcoI locus, NN genotype was not observed. Observed heterozygosity
was higher in MspI (0.22) than NcoI locus (0.03). MspI and NcoI loci were found
in Hardy-Weinberg equilibrium in Kivircik lambs raised in Kirklareli province. 

Keywords

Calpastatin native sheep Kivircik genetic variation

References

  • Aali, M., Moradi-Shahrbabak, M., Sadeghi, M., 2014. Detecting novel SNPs and breed-specific haplotypes at calpastatin gene in Iranian fat- and thin-tailed sheep breeds and their effects on protein structure. Gene 537, 132-139.
  • Asadi, N., Nanekarani, S., Khederzadeh, S., 2014. Genotypic frequency of calpastatin gene in lori sheep by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. African Journal of Biotechnology 13, 1952-1954.
  • Ata, N., Cemal, I., 2008. Calpastatin gene polymorphism in Çine Çapari and Karya sheep. Scientific Papers Series D Animal Science 56, 48-51.
  • Avanus, K., 2015. Genetic Variability of CAST Gene in Native Sheep Breeds of Turkey. Kafkas Üniversitesi Veteriner Fakültesi Dergisi 21(6), 789-794.
  • Azari, M.A., Dehnavi, E., Yousefi, S., Shahmohamadi, L., 2012. Polymorphism of calpastatin , calpain and myostatin genes in native Dalagh sheep in Iran. Slovak Journal of Animal Science 12, 1-6.
  • Byun, S.O., Zhou, H., Forrest, R.H.J., Frampton, C.M., Hickford, J.G.H., 2008. Association of the ovine calpastatin gene with birth weight and growth rate to weaning. Animal Genetics 39, 572-573.
  • Casas, E., White, S.N., Wheeler, T.L., Shackelford, S.D., Koohmaraie, R., Riley, D.G., Smith, T.P.L., 2006. Effects of calpastatin and μ-calpain markers in beef cattle on tenderness traits. Journal of Animal Science 84, 520-525.
  • Chung, H., Davis, M., 2012a. Effects of genetic variants for the calpastatin gene on calpastatin activity and meat tenderness in Hanwoo (Korean cattle). Meat Science 90; 711-714.
  • Chung, H., Davis, M., 2012b. PCR-RFLP of the ovine calpastatin gene and its association with growth. Asian Journal of Animal Science & Veterinary Advances 7, 641-652
  • Ciobanu, D.C., Bastiaansen, J.W.M., Lonergan, S.M., Thomsen, H., Dekkers, J.C.M., Rothschild, M.F., 2004. New alleles in calpastatin gene are associated with meat quality traits in pigs. Journal of Animal Science 82, 2829-2839.
  • Curi, R., Chardulo, L.L., Mason, M.C., Arrigoni, M.D.B., Silveira, C., Oliveira, H.N., 2009. Effect of single nucleotide polymorphisms of CAPN1 and CAST genes on meat traits in Nellore beef cattle (Bos indicus) and in their crosses with Bos taurus. Animal Genetics 40, 456-462.
  • Dagong, M.I., Sumantri, C., Noor, R.R., Herman, R., Yamin, M., 2011. Genetic polymorphisms of the coding region (exon 6) of calpastatin in Indonesian sheep. Media Peternakan 34, 190-195.
  • Dehnavi, E., Ahani Azari, M., Hasani, S., Nassiry, M.R., Mohajer, M., Khan, A.R., Shahmohamadi, L., Yousefi, S., 2012. Association between yearling weight and calpastatin and calpain loci polymorphism in Iranian Zel sheep. Iranian Journal of Applied & Animal Science 2, 131-135.
  • Djadid, N.D., Nikmard, M., Zakeri, S., Gholizadeh, S., 2011. Characterization of calpastatin gene in Iranian Afshari sheep. Iranian Journal of Biotechnology 9(2), 145-149.
  • Greguła-Kania, M., 2011. New allelic variant of the ovine calpastatin gene. African Journal of Biotechnology 10, 13082-13085.
  • Khan, S., Riaz, M.N., Ghaffar, A., Ullah, M.F., 2012. Calpastatin (CAST) gene polymorphism and its association with average daily weight gain in Balkhi and Kajli sheep and Beetal goat breeds. Pakistan Journal of Zoology 44(2), 377-382.
  • Khederzadeh, S., 2011. Polymorphism of calpastatin gene in crossbreed Dalagh sheep using PCR-RFLP. African Journal of Biotechnology 10, 10839-10841.
  • Mohammadi, M., Nasiri, M.T.B., Alami-Saeid, K., Fayazi, J., Mamoee, M., Sadr, A.S., 2008. Polymorphism of calpastatin gene in Arabic sheep using PCR- RFLP. African Journal of Biotechnology 7(15), 2682-2684.
  • Nanekarani, S., Khederzadeh, S., Kaftarkari, A.M., 2011. Genotypic frequency of calpastatin gene in Atabi sheep by PBR method. In: Proc. International Conference on Food Engineering and Biotechnology IPCBEE, Bangkok, Thailand, 9, 189-192.
  • Nassiry, M.R., Tahmoorespour, M., Javadmanesh, A., Soltani, M., Far, S.F., 2006. Calpastatin polymorphism and its association with daily gain in Kurdi sheep. Iranian Journal of Biotechnology 4(3), 188-192.
  • Page, B.T., Casas, E., Heaton, M.P., Cullen, N.G., Hyndman, D.L., Morris, C.A., Smith, T.P.L., 2002. Evaluation of single-nucleotide polymorphisms in CAPN1 for association with meat tenderness in cattle. Journal of Animal Science 80, 3077-3085.
  • Palmer, B.R., Roberts, N., Hickford, J.G., Bickerstaffe, R., 1998. PCR-RFLP for MspI and NcoI in the ovine calpastatin gene. Journal of Animal Science 76, 1499-1500.
  • Palmer, B.R., Su, H.Y., Roberts, N., Hickford, J.G., Bickerstaffe, R., 2000. Single nucleotide polymorphisms in an intron of the ovine calpastatin gene. Animal Biotechnology 11, 63-67.
  • Ranjbari, M., Hashemi, A., Mardani, K., Darvishzadeh, R., 2012. Allelic polymorphism of Makoei sheep calpastatin gene identified by polymerase chain reaction and single strand conformation polymorphism. Journal of Agricultural Science and Technology 14, 533-538.
  • Shahroudi, F.E., Nassiry, M.R., Valizadh, R., Heravi, A., 2006. Genetic poly-morphism at MTNR1A, CAST and CAPN loci in Iranian Karakul sheep. Iranian Journal of Biotechnology 4(2), 117-122.
  • Suleman, M., Khan, S.U., Riaz, M.N., Yousaf, M., Shah, A., Ishaq, R., 2012. Calpastatin (CAST) gene polymorphism in Kajli, Lohi and Thalli sheep breeds. African Journal of Biotechnology 4, 10655-10660.
  • Szkudlarek-Kowalczyk, M., Wiśniewska, E., Mroczkowski, S., 2011. Polymorphisms of calpastatin gene in sheep. Journal of Central European Agriculture 12, 425-432.
  • Yeh, F., Yang, R.C., Boyle, T., 2000. Popgene (v.1.32) Microsoft Windows-based freeware for Population Genetic Analysis: http://www.ualberta.ca/~fyeh/Pop32.exe (Accessed 07.09.2017)
  • Yılmaz, O., Cemal, İ., Karaca, O., Ata, N., 2014a. Association of Calpastatin (CAST) gene polymorphism with weaning weight and ultrasonic measurements of loin eye muscle in Kivircik lambs. Kafkas Universitesi Veteriner Fakültesi Dergisi 20, 675-680.
  • Yılmaz, O., Sezenler, T., Ata, N., Yaman, Y., Cemal, İ., Karaca, O., 2014b. Polymorphism of the ovine calpastatin gene in some Turkish sheep breeds. Turkish Journal of Veterinary & Animal Science 38, 354-357.
  • Zhou, H., Hickford, J.G.H., Gong, H., 2007. Polymorphism of the ovine calpastatin gene. Molecular and Cellular Probes 21, 242-244.
There are 31 citations in total.

Details

Primary Language English
Subjects Veterinary Surgery
Journal Section Original Article
Authors

Kozet Avanus

Publication Date January 16, 2018
Published in Issue Year 2018 Volume: 44 Issue: 1

Cite

APA Avanus, K. (2018). Determining Genetic Variation of Calpastatin Gene with MspI and NcoI Enzymes by Using PCR-RFLP Method in Kivircik Lambs. Acta Veterinaria Eurasia, 44(1), 39-43.
AMA Avanus K. Determining Genetic Variation of Calpastatin Gene with MspI and NcoI Enzymes by Using PCR-RFLP Method in Kivircik Lambs. Actavet. January 2018;44(1):39-43.
Chicago Avanus, Kozet. “Determining Genetic Variation of Calpastatin Gene With MspI and NcoI Enzymes by Using PCR-RFLP Method in Kivircik Lambs”. Acta Veterinaria Eurasia 44, no. 1 (January 2018): 39-43.
EndNote Avanus K (January 1, 2018) Determining Genetic Variation of Calpastatin Gene with MspI and NcoI Enzymes by Using PCR-RFLP Method in Kivircik Lambs. Acta Veterinaria Eurasia 44 1 39–43.
IEEE K. Avanus, “Determining Genetic Variation of Calpastatin Gene with MspI and NcoI Enzymes by Using PCR-RFLP Method in Kivircik Lambs”, Actavet, vol. 44, no. 1, pp. 39–43, 2018.
ISNAD Avanus, Kozet. “Determining Genetic Variation of Calpastatin Gene With MspI and NcoI Enzymes by Using PCR-RFLP Method in Kivircik Lambs”. Acta Veterinaria Eurasia 44/1 (January 2018), 39-43.
JAMA Avanus K. Determining Genetic Variation of Calpastatin Gene with MspI and NcoI Enzymes by Using PCR-RFLP Method in Kivircik Lambs. Actavet. 2018;44:39–43.
MLA Avanus, Kozet. “Determining Genetic Variation of Calpastatin Gene With MspI and NcoI Enzymes by Using PCR-RFLP Method in Kivircik Lambs”. Acta Veterinaria Eurasia, vol. 44, no. 1, 2018, pp. 39-43.
Vancouver Avanus K. Determining Genetic Variation of Calpastatin Gene with MspI and NcoI Enzymes by Using PCR-RFLP Method in Kivircik Lambs. Actavet. 2018;44(1):39-43.
 Download Cover Image