Molecular Biology Reports

, Volume 36, Issue 3, pp 455–460 | Cite as

Molecular cloning and expression analyses of a novel swine gene-ARF4

  • G. Y. Liu
  • Y. Z. Xiong


The mRNA differential display technique was performed to investigate the differences of gene expression in the longissimus muscle tissues from Meishan and Large White pigs. One novel gene that was differentially expressed was identified through semi-quantitative RT-PCR and the cDNA complete sequence was then obtained using the rapid amplification of cDNA ends (RACE) method. The nucleotide sequence of the gene is not homologous to any of the known porcine genes. The sequence prediction analysis revealed that the open reading frame of this gene encodes a protein of 180 amino acids that contains the putative conserved domain of ADP-ribosylation factor (ARF) which has high homology with the ADP-ribosylation factor 4 (ARF4) of six species—bovine (98%), human and orangutan (96%), African clawed frog (96%), mouse and rat (98%)—so that it can be defined as swine ADP-ribosylation factor 4 (ARF4). This novel porcine gene was finally assigned to GeneID:595108. The phylogenetic tree analysis revealed that the swine ARF4 has a closer genetic relationship with the rat and mouse ARF4 than with those of human and African clawed frog. The tissue expression analysis indicated that the swine ARF4 gene is over expressed in muscle, fat, heart, spleen, liver, and ovary and moderately expressed in lung and kidney but weakly expressed in small intestine. Our experiment is the first to establish the primary foundation for further research on the swine ARF4 gene.


Pig mRNA differential display RACE 


  1. 1.
    Liang P, Pardee AB (1992) Differential display of eukaryotic messenger RNA by means of the polymerase chain reaction. Science 257(5072):967–971PubMedCrossRefGoogle Scholar
  2. 2.
    Liang P, Averboukh L, Pardee AB (1993) Distribution and cloning of eukaryotic mRNAs by means of differential display: refinements and optimization. Nucleic Acids Res 21(14):3269–3275PubMedCrossRefGoogle Scholar
  3. 3.
    Yamazaki M, Saito K (2002) Differential display analysis of gene expression in plants. Cell Mol Life Sci 59(8):1246–1255PubMedCrossRefGoogle Scholar
  4. 4.
    Pan PW, Zhao SH, Yu M, Xiong TA, Li K (2003) Identification of differentially expressed genes in the Longissimus dorsi tissue between Duroc and Erhualian pigs by mRNA differential display. Asian-Aust J Sci 16(7):1066–1070Google Scholar
  5. 5.
    Liu GY, Xiong YZ, Deng CY, Zuo B, Zhang JH (2004) Comparison of gene expression patterns in Longissimus dorsi of pigs between the high-parent heterosis cross combination Landrace × Large White and the mid-parent heterosis cross combination Large White × Meishan. Asian-Aust J Anim Sci 17(9):1192–1196Google Scholar
  6. 6.
    Liu GY, Xiong YZ, Deng CY (2005) Isolation, identification of differentially expressed sequence tags in the backfat tissue from Meishan, Large White and Meishan × Large White cross pigs. Agric Sci China 4(1):101–105Google Scholar
  7. 7.
    Daigo Y, Takayama I, Ponder BA, Caldas C, Ward SM, Sanders KM, Fujino MA (2003) Differential gene expression in the murine gastric fundus lacking interstitial cells of Cajal. BMC Gastroenterol 3(1):14PubMedCrossRefGoogle Scholar
  8. 8.
    Fehr JE, Trotter GW, Oxford JT, Hart DA (2000) Comparison of Northern blot hybridization and a reverse transcriptase-polymerase chain reaction technique for measurement of mRNA expression of metalloproteinases and matrix components in articular cartilage and synovial membrane from horses with osteoarthritis. Am J Vet Res 61(8):900–905PubMedCrossRefGoogle Scholar
  9. 9.
    Liu YG, Xiong YZ, Deng CY (2005) Isolation, sequence analysis and expression profile of a novel swine gene differentially expressed in the Longissimus dorsi muscle tissues from Landrace × Large White cross-combination. Acta Biochimica et Biophysica Sinica 37(3):186–191PubMedCrossRefGoogle Scholar
  10. 10.
    Kahn RA, Kern FG, Clark J, Gelmann EP, Rulka C (1991) Human ADP-ribosylation factors. A functionally conserved family of GTP-binding proteins. J Biol Chem 266(4):2606–2614PubMedGoogle Scholar
  11. 11.
    Mandiyan V, Andreev J, Schlessinger J, Hubbard SR (1999) Crystal structure of the ARF-GAP domain and ankyrin repeats of PYK2-associated protein beta. EMBO J 18(24):6890–6898PubMedCrossRefGoogle Scholar
  12. 12.
    Shome K, Nie Y, Romero G (1998) ADP-ribosylation factor proteins mediate agonist-induced activation of phospholipase D. J Biol Chem 273(46):30836–30841PubMedCrossRefGoogle Scholar
  13. 13.
    Moss J, Vaughan M (1995) Structure and function of ARF proteins: activators and cholera toxin and critical components of intracellular vesicular transport processes. J Biol Chem 270:12327–12330PubMedCrossRefGoogle Scholar
  14. 14.
    Greasley SE, Jhoti H, Teahan C, Solari R, Fensome A, Thomas GM, Cockcroft S, Bax B (1995) The structure of rat ADP-ribosylation factor-1 (ARF-1) complexed to GDP determined from two different crystal forms. Nat Struct Biol 2(9):797–806PubMedCrossRefGoogle Scholar
  15. 15.
    Amor JC, Harrison DH, Kahn RA, Ringe D (1994) Structure of the human ADP-ribosylation factor 1 complexed with GDP. Nature 372(6507):704–708PubMedCrossRefGoogle Scholar
  16. 16.
    Kim SW, Hayashi M, Lo JF, Yang Y, Yoo JS, Lee JD (2003) ADP-ribosylation factor 4 small GTPase mediates epidermal growth factor receptor-dependent phospholipase D2 activation. J Biol Chem 278(4):2661–2668PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2007

Authors and Affiliations

  1. 1.Key Laboratory of Animal Nutrition and Feed of Yunnan ProvinceYunnan Agricultural UniversityKunmingChina
  2. 2.Key Laboratory of Swine Genetics and Breeding, Ministry of AgricultureHuazhong Agricultural UniversityWuhanChina

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