مطالعۀ چندشکلی ژن ACACA و ارتباط آن با صفات رشد و تولید شیر در بزهای نژاد مهابادی با روش PCR-SSCP

نوع مقاله : مقاله پژوهشی

نویسندگان

1 دانشجوی سابق کارشناسی ارشد رشتۀ ژنتیک و اصلاح نژاد دام، گروه علوم دامی پردیس کشاورزی و منابع طبیعی دانشگاه تهران، کرج

2 دانشیار، گروه علوم دامی پردیس کشاورزی و منابع طبیعی دانشگاه تهران، کرج

3 استاد، گروه علوم دامی پردیس کشاورزی و منابع طبیعی دانشگاه تهران، کرج

چکیده

این پژوهش با هدف مطالعۀ چندشکلی ژن ACACA و ارتباط آن با صفات شیر در بزهای نژاد مهابادی انجام گرفت. به این منظور برای 150 رأس بز شیری نژاد مهابادی، خون‌گیری و استخراج DNA از خون به روش نمکی بهینه‌یافته صورت گرفت. قطعۀ اینترون 44 ژن ACACA، به کمک واکنش‌های زنجیره‌ای پلیمراز تکثیر شد. برای بررسی چندشکلی قطعات تکثیرشده، از روش چندشکلی فضایی تک‌رشته‌ای DNA (SSCP) استفاده شد. در نهایت هفت الگوی متفاوت به دست آمد که با هم‌ردیفی و مقایسۀ توالی‌ها با یکدیگر و با توالی ژن ACACA بز با شمارۀ دسترسی NC_022311، در مجموع هفت جهش در جمعیت بز مهابادی در موقعیت‌های 154503(تغییر باز T به G)، 154518(تغییر باز G به A)، 154577(تغییر باز G به A)، 154640(تغییر باز T به C)، 154712(تغییر باز C به A)، 154928(تغییر باز T به C)، 154956(تغییر باز C به T) مشاهده شد. ارتباط هر  SNP با صفات تولید شیر (مقدار تولید، درصد چربی و پروتئین و تعداد سلول‌های بدنی) با نرم‌افزار SAS 9.1 بررسی شد و نتایج حاکی از اثر معنا‌دار SNP دوم (05/0P=)، سوم (05/0P=) و پنجم (01/0>P) بر تولید شیر و SNP اول (017/0P=)، ششم (017/0P=) و هفتم (01/0>P) بر درصد چربی شیر بود.

کلیدواژه‌ها


عنوان مقاله [English]

Polymorphism in ACACA gene and its association with milk and growth traits in Mahabadi goats using the PCR-SSCP technique

نویسندگان [English]

  • Reza Faraji 1
  • Mostafa Sadeghi 2
  • Mohammad Moradi Shahrbabak 3
1 Former M. Sc. Student, Department of Animal Science, University College of Agriculture & Natural Resources, University of Tehran, Karaj, Iran
2 Associate Professor, Department of Animal Science, University College of Agriculture & Natural Resources, University of Tehran, Karaj, Iran
3 Professor, Department of Animal Science, University College of Agriculture & Natural Resources, University of Tehran, Karaj, Iran
چکیده [English]

This research was conducted to study polymorphism of intron 44 of ACACA gene and its association with milk traits in Mahabadi goats. Blood samples were taken from 81 Mahabadi goats. DNA was extracted from blood using salting out method. 897 base pair fragment of intron 44 ACACA gene was amplified using polymerase chain reaction. SSCP method was used to investigate the amplified DNA fragment polymorphism. Amplified products were sequenced in both directions. Finally, seven different patterns were obtained and sequences compared with each other also with Capra hircus ACACA gene sequence on NCBI (NC_022311). Seven novel SNPs were identified which were NC_022311: nucleotide 154503(T/G), 154518(A/G), 154577(A/G), 154640(C/T), 154712(A/C), 154928(C/T), 154956(C/T). Then a mixed animal model was used by SAS9.1 analysis system to evaluate the association of these SNPs with milk traits. In this model, variables of age of dam, genotypes and mount of recording were considered as fixed effects. There were significant association of SNP2 (P=0.05), SNP3 (P=0.05) and SNP5 (P<0.01) with milk yield. SNPs 1 (P=0.017), 6 (P=0.017) and 7 (P<0.01) had also significant effect on milk fat-percentage.

کلیدواژه‌ها [English]

  • ACACA gene
  • Mahabadi goats
  • milk traits
  • PCR-SSCP
  • Polymorphism
  1. Badaoui, B., Serradilla, J., Tomas, A., Urrutia, B., Ares, J., Carrizosa, J., Sanchez, A., Jordana, J. & Amills, M. (2007). Goat acetyl-coenzyme A carboxylase α: molecular characterization, polymorphism, and association with milk traits. Journal of Dairy Science, 90(2), 1039-1043.
  2. Barber, M., Pooley, L. & Travers, M. (2001). Developmental regulation of alternatively spliced acetyl-CoA carboxylase-alpha mRNAs encoding isozymes with or without an eight amino acid domain upstream of the Ser-1200 phosphorylation motif in the mammary gland. Journal of Molecular Endocrinology, 27(3), 349-356.
  3. Barber, M.C. & Travers, M.T. (1998). Elucidation of a promoter activity that directs the expression of acetyl-CoA carboxylase alpha with an alternative N-terminus in a tissue-restricted fashion. Biochemical Journal, 333(Pt 1), 17.
  4. Crepaldi, B., Nicoloso, L., Coizet, B., Milanesi, E., Pagnacco, G., Fresi, P. & Dimauro, C. (2013). Associations of acetyl-coenzyme A carboxylase α, stearoyl-coenzyme A desaturase, and lipoprotein lipase genes with dairy traits in Alpine goats. Journal of Dairy Science, 96, 1856-1864.
  5. Dong, Y., Xie, M., Jiang, Y., Xiao, N., Du, X., Zhang, W., Tosser-Klopp, G., Wang, J., Yang, S., Liang, J., Chen, W., Chen, J., Zeng, P., Hou, Y., Bian, C., Pan, S., Li, Y., Liu, X., Wang, W., Servin, B., Sayre, B., Zhu, B., Sweeney, D., Moore, R., Nie, W., Shen, Y., Zhao, R., Zhang, G., Li, J., Faraut, T., Womack, J., Zhang, Y., Kijas, J., Cockett, N., Xu, X., Zhao, S., Wang, J. & Wang, W. (2013). Sequencing and automated whole-genome optical mapping of the genome of a domestic goat (Capra hircus). Nature Biotechnology, 31(2), 135-41.
  6. Federica, S., Francesco, N., Giovanna, D M., Carmela, SM., Gennaro C., Carmela, T. & Bianca, M. (2009). Identification of novel single nucleotide polymorphisms in promoter III of the acetyl-CoA carboxylase-{alpha} gene in goats affecting milk production traits. Journal of Hered, 100(3), 386-9.
  7. Gallardo, D., Quintanilla, R., Varona, L., Díaz, I., Ramirez, O., Pena, R. & Amills, M. (2009). Polymorphism of the pig acetyl‐coenzyme A carboxylase α gene is associated with fatty acid composition in a Duroc commercial line. Animal Genetics, 40(4), 410-417.
  8. García-Fernández, M., Gutiérrez-Gil, B., Garcia-Gámez, E. & Arranz, J.J. (2010). Identification of single nucleotide polymorphisms in the ovine acetyl-CoA carboxylase-alpha gene. Small Ruminant Research, 90(1), 34-40.
  9. Ibeagha-Awemu, E.M., Kgwatalala, P. & Zhao, X. (2008). A critical analysis of production-associated DNA polymorphisms in the genes of cattle, goat, sheep, and pig. Mammalian Genome, 19(9), 591-617.
  10. Matsumoto, H., Sasaki, K., Bessho, T., Kobayashi, E., Abe, T., Sasazaki, S., Oyama, K. & Mannen, H. (2012). The SNPs in the ACACA gene are effective on fatty acid composition in holstein milk. Molecular Biology Reports: 1-8.
  11. Moioli, B., D’Andrea, M. & Pilla, F. (2007). Candidate genes affecting sheep and goat milk quality. Small Ruminant Research, 68(1), 179-192.
  12. Moioli, B., Napolitano, F., Orrù, L. & Catillo, G. (2005). Single nucleotide polymorphism detection in promoter I of the acetyl-CoA carboxylase-α gene in sheep. Small Ruminant Research, 59(1), 49-53.
  13. Moioli, B., Napolitano, F., Orru, L. & Catillo, G. (2005). Single nucleotide polymorphism detection in promoter III of the acetyl‐CoA carboxylase‐α gene in sheep. Journal of Animal Breeding and Genetics, 122(6), 418-420.
  14. Shin, S.C., Heo, J. & Chung, E.R. (2011). Effect of Single Nucleotide Polymorphisms of Acetyl-CoA Carboxylase α (ACACA) Gene on Carcass Traits in Hanwoo (Korean Cattle).
  15. Tong, L. (2005). Acetyl-coenzyme A carboxylase: crucial metabolic enzyme and attractive target for drug discovery. Cellular and Molecular Life Sciences, 62(16), 1784-1803.
  16. Travers, M. & Barber, M. (2001). Acetyl-CoA carboxylase-α: gene structure-function relationships. Journal of Animal Science, 79(Suppl E), E136-E143.
  17. Volpe, J. & Marasa, J. (2006). Regulation of palmitic acid synthesis in cultured glial cells: Effects of lipid on Fatty acid synthetase, acetyl‐CoA carboxylase, fatty acid and sterol synthesis1. Journal of Neurochemistry, 25(3), 333-340.
  18. Zhang, S., Knight, T., Reecy, J., Wheeler, T., Shackelford, S., Cundiff, L. & Beitz, D. (2009). Associations of polymorphisms in the promoter I of bovine acetyl‐CoA carboxylase‐α gene with beef fatty acid composition. Animal Genetics, 41(4), 417-420.
  19. Zhang, S., Knight, T.J., Reecy, J.M., Wheeler, T.L., Shackelford, S.D., Cundiff, L.V. & Beitz, D.C. (2008). Association of polymorphism in the promoter I of bovine Acetyl-CoA Carboxylase- alpha gene with beef lipid content. Genetcs, 3, 123-132.