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Identification of Meat Species Using PCR-RFLP Marker of Cytochrome b Gene 원문보기

Korean journal for food science of animal resources = 한국축산식품학회지, v.26 no.3, 2006년, pp.375 - 379  

Shin, Sung-Chul (Division of Animal Science and Resources, College of Life Science and Natural Resources, Sangji University) ,  Chung, Ku-Young (Division of Animal Science and Resources, College of Life Science and Natural Resources, Sangji University) ,  Chung, Eui-Ryong (Division of Animal Science and Resources, College of Life Science and Natural Resources, Sangji University)

Abstract ▼ AI-Helper 아이콘AI-Helper

Food labeling regulations require that the meat species in various meat products are accurately declared to the consumer. Substitution or adulteration of costly meat with a cheaper one is one of the most common problems in the meat industry. In this study, PCR-restriction fragment length polymorphis...

주제어

#cytochrome b gene   #PCR-RFLP   #meat species identification  

AI 본문요약
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제안 방법

  • 1989). The primer sequences were as follows: CYTb F 5'- CCA TCC AAC ATC TCA GCA TGA TGA AA-3' and CYTb R 5'- GCC CCT CAG AAT GAT ATT TGT CCT CA-3\ PCR amplifications were carried out in a final volume of 20 "L containing 50 ng template DNA, 0.1 "M of each primer, 250 #M of each dNTP, 1.0 U of Taq DNA polymerase and 2 "L of 10 X PCR buffer. The PCR reaction was performed with a Perkin-Elmer (GeneAmp PCR system 9700, PE Biosystem, Foster City, CA) thermal cycler according to the following PCR step-cycle program: pre-denaturation of 94°C for 1 min, followed by 30 cycles of denaturation at 94 °C fbr 20 s, annealing at 55 °C for 30 s and extension at 72 °C for 40 s, and final extension at 72 °C for 2min
  • Based on analysis of the restriction map of sequences Hae HI and Hirfi enzymes were chosen for RFLP analysis, so as to identify and differentiate meat species. The PCR products digested with Hae]R and Hinft enzymes showed species­ specific RFLP patterns according to their restriction profiles, respectively, as shown in Fig.

대상 데이터

  • , 1995). Sequence comparison of mt cyt b gene available in the GenBank/EMBL database fbr several animal species permitted the design of 2 conserved primers, CYTZ? F and CYTb R, which amplified a DNA fragment of approximately 359 bp from nine meat species examined in this study. Nucleotide substitutions within and between mt cyt b gene sequences are sufficient for species identification of different biological samples (Prakash et al.
  • Nine meat species from beef (Bos taurus)^ pork (Sus scro- Ja), horse (Equus caballus), goat (Capra hircus), mutton (Ovis aries), deer (Cervus nippon), chicken (Gallus gallus), turkey (Meleagris gallopavo) and duck (Anas boschas) were used in this study. All meat samples were obtained from commercial sources.

이론/모형

  • In this study, the mt cyt b gene was used as a molecukr marker for the identification of meat species using PCR-RFLP method. To amplify a DNA fragment from the mt cyt b gene of the six mammalian species (cattle, pig, horse, dear, sheep and goat) and three poultry species (chicken, turkey and duck), a specific universal primer pair was designed and synthesized based on sequences available in the GenBanl-7 EMBL database for several animal species.
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참고문헌 (19)

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  5. Dooley, J. J., Paine, K. E., Garrett, S. D., and Brown, H. B. (2004) Detection of meat species using TaqMan real-time PCR assays. Meat Sci. 68, 431-438 

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  6. Girish, P. S., Anjaneyulu, A. S. R., Viswas, K. N., Shivakumar, B. M., Anand, M., Patel, M., and Sharma, B. (2005) Meat species identification by polymerase chain reation-restriction fragment length polymorphism (PCRRFLP) of mitochondrial 12S rRNA gene. Meat Sci. 70, 107-112 

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  7. Hird, H., Gooder, R., and Hill, M. (2003) Rapid diction of chicken and turkey in heated meat products using the polymerase chain reaction followed by amplicon visualization with vistra green. Meat Sci. 65, 1117-1123 

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  8. Kocher, T. D., Thomas, W. K., Mayer, A., Edwards, S. V., Paabo, S., Villablanca, F. X., and Wilson, A. C. (1989) Dynamics of mitochondrial DNA evolution in animals: Amplification and sequencing with conserved primers. Proc. Natl. Acad. Sci. USA 86, 6196-6200 

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  12. Meyer, R, Hofelein, C., and Luthy, J. (1994) Detection of pork in heated meat products by the polymerase chain reaction. J. Assoc. Off. 77, 617-622 

  13. Meyer, R., Hoefelein, C., Luethy, J., and Candrian, U. (1995) Polymerase chain reaction-restriction fragment length polymorphism analysis: a simple method for species identification in food. J. AOAC Int. 78, 1542-1552 

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  15. Partis, L., Croan, D., Guo, Z., Clark, R, Coldham, T., and Murby, J. (2000) Evaluation of a DNA fingerprinting method for determining the species origin of meats. Meat Sci. 54, 369-376 

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  16. Prakash, P. S., Ghumatkar, M. S., Nandode, S. V., Yogesh, S. K., and Shouche, H. Y. (2000) Mitochondrial 12S rRNA sequence analysis in wild life forensics. Current Sci. 78, 1542-1551 

  17. Rodriquez, M. A., Garcia, T., Gonzalez, I., Asensio, L., Mayoral, B., Lopez-Calleja, I. Hernandez, P. E., and Martin, R. (2003) Development of polymerase chain reaction assay for species identification of goose and mule duck in foie gras products. Meat Sci. 65, 1257-1263 

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  18. Rodriguez, M. A., Garcia, T., Gonzalez, I., Hernandez, P. E., and Martin, R. (2005) TaqMan real-time PCR for the detection and quantitation of pork in meat mixtures. Meat Sci. 70, 113-120 

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  19. Sawyer, J., Wood, C., Shanahan, D., Gout, S., and McDowell, D. (2003) Real-time PCR for quantitative meat species testing. Food Cont. 14, 579-583 

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