Investigation of Iranian native Sarabi Cows for Complex Vertebral Malformation (CVM) and Deficiency in Uridine Mono Phosphate Syntase Enzyme (DUMPS) Genetic Disorders by Using PCR-SSCP and PCR-RFLP Technique

Document Type : Research Paper

Authors

Abstract

Evaluating and detection of genetic disorders is very important in both human and animal communities. Blood samples were prepared from 162 cows from Sarabi cows’ research station in Sarab, East Azerbaijan province, Iran. DNA extraction was done based on guanidine thiocyanate – silica gel method. The aim of this study was to identify the CVM and DUMPS carriers in Sarabi native cows of Iran. Standard polymerase chain reaction was applied to amplify a 177 bp and a 108 bp fragments from exon 4 in SLAC35A3 gene and exon 5 in UMPs gene using specific primers pair. The SLC35A3 gene PCR products were analyzed using single stranded conformation polymorphism (SSCP) to reveal their mutations on polyacrylamide gel. In order to identify DUMPS carrier animals, a RFLP assay was conducted and the UMPs PCR products were digested by AvaI restriction enzyme. Results indicated that no G→T mutation at position 559 in exon 4 of SLAC35A3 gene and C→T mutation at position 405 in exon 5 of UMPs gene in examined cows respectively and subsequently no carrier was identified.

 

Keywords

References

  1. Agerholm JS, Bendixen C, Andersen O, Arnbjerg J (2000) Complex vertebral malformations in Holstein calves. Report LK-564. National Committee on Danish Cattle Husbandry, Aarhus, Denmark.
  2. Agerholm JS, Bendixen C, Andersen O, Arnbjerg J (2001) Complex vertebral malformation in Holstein calves. Journal of Veterinary Diagnostic Investigation 13:283–289.
  3. Agerholm JS, Willadsen CM, Nielsen TK, Giese SB, Holm E, Jensen L, Agger JF (1997) Diagnostic studies of abortion in Danish dairy herds. Journal of Veterinary Medicine A 44: 551-558.
  4. Akyuz B, Ertugrul O (2008) Detection of deficiency of Uridine Monophosphate Synthase (DUMPS) in Holstein and native cattle in Turkey. Journal of Ankara University Faculty of Veterinary Medicine 55: 57-60.
  5. Duncan RB, Colin Jr, Carrig B, Agerholm JS, Bendixen C (2001) Complex vertebral malformation in a Holstein calf: report of a case in the USA. Journal of Veterinary Diagnostic Investigation 13:333–336.
  6. Freeman AR, Meghen CM, Machugh DE, Loftus RT, Achukwi MD, Bado A, Sauveroche B, Bradley DG (2004) Admixture and diversity in West African cattle populations. Molecular Ecology 13: 3477-3487.
  7. Harlizius B, Schrober S, Tammen I, Simon D (1996) Isolation of the bovine uridine monophosphate synthase gene to identify the molecular basis of DUMPS in cattle. Journal of Animal Breeding and Genetics 133: 303-309.
  8. Herzog A (1992) Genetic defects in cattle and the possibilities of control. Wiener Tierarztliche Monatsschrift 79: 142-148
  9. Ishida N, Kawakita M (2004) Molecular physiology and pathology of the nucleotide sugar transporter family (SLC35). Pflügers Archiv European Journal of Physiology 447: 768-775
  10. Ishida N, Kuba T, Aoki K, Miyatake S, Kawakita M, Sanai Y (2005) Identification and characterization of human Golgi nucleotide sugar transporter SLC35D2, a novel member of the SLC35 nucleotide sugar transporter family. Genomics 85: 106-116.
  11. Kaminski S, Grzybowski G, Prusak B, Rusc A (2005) No incidence of DUMPS carriers in Polish dairy cattle. Journal of Applied Genetics 46 (4): 395-397.
  12. Kanae Y, Endoh D, Nagahata H, Hayashi, M (2005) A method for detecting complex vertebral malformation in Holstein calves using polymerase chain reaction–primer introduced restriction analysis. Journal of Veterinary Diagnostic Investigation 17: 258-262.
  13. Muraoka M, Kawakita M, Ishida, N (2001) Molecular characterization of human UDP-glucuronic acid/UDP-N-acetylgalactosamine transporter, a novel nucleotide sugar transporter with dual substrate specificity. FEBS Letters 495: 87-93.
  14. Nagahata H, Oota H, Nitanai A, Oikawa S., Higuchi H., Nakade T, Kurosawa T, Morita M, Ogawa H. (2002). Complex Vertebral Malformation in a Stillborn Holstein Calf in Japan. Journal of Veterinary Medical Science. 64(12): 1107-1112.
  15. Patel RK, Singh KM., Soni KJ, Chauhan JB, Sambasiva Rao KRS (2006) Lack of carriers of citrullinaemia and Dumps in Indian Holstein cattle. Journal of Applied Genetics 47(3): 239-242.
  16. Rusc A, Kaminski S (2007). Prevalence of complex vertebral malformation carriers among Polish Holstein-Friesian bulls. Journal of Applied Genetics 48(3): 247-252.
  17. Schwenger B, Schober S, Simon D (1993). DUMPS cattle carry a point mutation in the Uridine Monophosphate synthase gene. Genomics 16: 241-244.
  18. Shanks RD, Robinson JL (1990) Deficiency of uridine monophosphate synthase among Holstein cattle. Cornell Veterinarian 80: 119-122.
  19. Thomsen B, Horn P, Panitz F, Bendixen E, Petersen A. H, Holm L (2006) A missense mutation in the bovine SLC35A3 gene, encoding a UDP-N-acetylglucosamine transporter, causes complex vertebral malformation. Genome Research 16: 97-105.
Agricultural Biotechnology Journal

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Available Online from 06 March 2011

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