Asian-Australasian Journal of Animal Sciences

Asian Australasian Association of Animal Production Societies (아세아태평양축산학회)
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Molecular Cloning, Characterization, and Expression Analysis of Chicken Δ-6 Desaturase

  • Kang, Xiangtao (College of Animal Science and Technology, Gansu Agricultural University) ;
  • Bai, Yichun (College of Animal Husbandry and Veterinary Science, Henan Agricultural University) ;
  • Sun, Guirong (College of Animal Husbandry and Veterinary Science, Henan Agricultural University) ;
  • Huang, Yanqun (College of Animal Husbandry and Veterinary Science, Henan Agricultural University) ;
  • Chen, Qixin (College of Animal Husbandry and Veterinary Science, Henan Agricultural University) ;
  • Han, Ruili (College of Animal Husbandry and Veterinary Science, Henan Agricultural University) ;
  • Li, Guoxi (College of Animal Husbandry and Veterinary Science, Henan Agricultural University) ;
  • Li, Fadi (College of Animal Science and Technology, Gansu Agricultural University)
  • Received : 2009.01.04
  • Accepted : 2009.05.17
  • Published : 2010.01.01
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Abstract

Long-chain polyunsaturated fatty acids (LC-PUFA) promote the development of brain and vision of the fetus, relieve inflammation, inhibit oral dysplasia of rumor cell, decrease the incidence of cardiovascular disease and regulate arrhythmia. ${\Delta}-6$ desaturase is the rate-limited enzyme in the desaturation process. This study reports the cloning, characterization and tissue expression of a ${\Delta}-6$ desaturase gene in the chicken. PCR primers were designed based on the predicted sequence of chicken ${\Delta}-6$ desaturase (accession number: XM421053) and used to isolate a cDNA fragment of 1,323 bp from chicken liver. Based on the 1,323 bp fragment an EST (BI390105) was obtained by BLAST. The EST and 5'nd of the 1,323 bp fragment were partially overlapped. Gene specific primers derived from the EST were used for amplification of the 5'nd. Another gene-specific primer derived from the 1,323 bp fragment was used for amplification of the 3'nd by 3'ACE. Then the three overlapping cDNA sequences obtained were assembled with DNAMAN software and a full-length ${\Delta}-6$ desaturase of 2,153 bp was obtained. The full-length cDNA contained an ORF of 1,335 bp with a 5'ntranslated region of 147 nucleotides followed by an ATG initiation codon. Stop codon TGA was at position 1,481-1,483 bp. The deduced amino acids shared an homology above 77% with bovine, mice, orangutan, rat and human. The protein sequence had three histidine-rich regions HDFGH (HisI region), HFQHH (HisII region) and HH (HisIII region), a cytochrome $b_{5}$-like domain containing a heme-binding motif and two transmembrane domains. Sequence analysis of the chicken genomic DNA revealed that the coding sequence of chicken ${\Delta}-6$ desaturase included 12 exons and 11 introns. Semi-quantitative RT-PCR showed that the ${\Delta}-6$ desaturase expression levels were in turn liver, spleen, pancreas, lung, breast muscle, heart, and abdominal fat. The expression of ${\Delta}-6$ desaturase in liver was significantly higher than that in breast muscle (p<0.01). The expression of ${\Delta}-6$ desaturase in lung was significantly higher than that in abdominal fat (p<0.01). This is the first clone of chicken ${\Delta}-6$ desaturase.

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References

  1. Andrea, D. S., H. Guillou, S. Jan, D. Catheline, J. N. Thibault, I. Bouriel, V. Rioux and P. Legrand. 2002. The same rat delta6-desaturase not only acts on 18- but also on 24- carbon fatty acids in very-long-chain polyunsaturated fatty acid biosynthesis. Biochem. J. 364(1):49-55
  2. Farrell, P. M., G. R. Gutcher, M. Palta and D. DeMet. 1988. Essential fatty acid deficiency in premature infant. Am. J. Clin. Nutr. 48(2):220-229
  3. Guillou, H., V. Rioux, D. Catheline, J. N. Thibault, I. Bouriel, S. D. Jan, S. Andrea and P. Legrand. 2003. Conversion of hexadecanoic acid to hexadecenoic acid by rat Delta 6-desaturase. J. Lipid Res. 44(3):450-454 https://doi.org/10.1194/jlr.C200019-JLR200
  4. Khang, N. T., D. G. Jennen, E. Tholen, D. Tesfaye, L. Mennicken, M. Hoelker, K. Schellander, S. Ponsuksili, E. Murani and K. Wimmers. 2007. Association of the FADS2 gene with omega-6 and omega-3 PUFA concentration in the egg yolk of Japanese quail. Anim. Biotechnol. 18(3):189-201 https://doi.org/10.1080/10495390701201390
  5. Marquardt, A., H. Stohr, K. White and B. H. Weber. 2000. cDNA Cloning, genomic structure, and chromos- omal localization of three members of the human fatty acid desaturase family. Genomics 66(2):175-183 https://doi.org/10.1006/geno.2000.6196
  6. Narce, M., P. Asdrubla, M. C. Delachambre, J. Gresti and J. P. Poisson. 1995. Influence of spontaneous hypertension on n-3 delta-6-desaturase activity and fatty acid composition of rat hepatocytes. Mol. Cell Biochem. 152(1):7-12
  7. Okayasu, T., M. Nagao, T. Ishibashi and Y. Imai. 1981. Purification and partial characterization of linoleoyl CoA desaturase from rat liver microsomes. Arch Biochem Biophys. 206(1):21-28 https://doi.org/10.1016/0003-9861(81)90061-8
  8. Reddy, A. S., M. L. Nuccio, L. M. Rross and T. L. Thomas. 1993. Isolation of a delta6-desaturase gene from the cyanobacterium Synechocystis sp. strain PCC 6803 by gain-of-function expression in Anabaena sp. strain PCC 7120. Plant Mol. Biol. 22(2):293-300 https://doi.org/10.1007/BF00014936
  9. Seiliez, I. S., S. Panserat, P. Kaushik and Bergot. 2001. Cloning, tissue distribution and nutritional regulation of a $\Delta$6-desaturase-like enzyme in rainbow trout. Comp. Biochem. Physiol. B Biochem. Mol. Biol. 130(1):83-93 https://doi.org/10.1016/S1096-4959(01)00410-9
  10. Zheng, X., I. Seiliez, N. Hastings, D. R. Tocher, S. Panserat, C. A. Dickson, P. Bergot and A. J. Teale. 2004. Characterization and comparison of fatty acyl delta6 desaturase cDNAs from freshwater and marine teleost fish species. Comp. Biochem. Physiol. B Biochem. Mol. Biol. 139 (2):269-279 https://doi.org/10.1016/j.cbpc.2004.08.003
  11. Zheng, X., D. R. Tocher, C. A. Dickson, J. G. Bell and A. J. Teale. 2005. Highly unsaturated fatty acid synthesis in vertebrates: new insights with the cloning and characterization of a delta6 desaturase of Atlantic salmon. Lipids 40(1):13-24 https://doi.org/10.1007/s11745-005-1355-7

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