Slc7a11 belongs to solute transporter gene family, encoding cystine/glutamate transporter xCT. It regulates switching between eumelanin and pheomelanin synthesis. In the present study, Real-time PCR was used to detect the mRNA expression levels of Slc7a11 in the skin of Kazakh lambs with different coat colors (black, brown and white), and then the prokaryotic expression plasmid PET-32a-sxCT was constructed to induce the expression of fusion protein. The target protein was purified by Ni-NTA affinity chromatographic separation, and then was used to immunize rabbit in order to produce rabbit anti-sxCT polyclonal antibody. Finally, the expression levels of sxCT were detected in the skin of Kazakh lambs with different hair colors by Western blotting analysis. Results showed that the mRNA expression levels of Slc7a11 differed significantly in the skin of Kazakh lambs with different coat colors, with the highest level in brown coat color, followed by the black, and then the white. The sxCT protein was also detected in the skin of different coat colors by polyclonal antibody, with the highest level in brown coat color, followed by the black, and then the white. It is, therefore, concluded that slc7a11 gene might be associated with the phenotype of coat color in Kazakh sheep.
LI Hong-Tao, HE Xin, ZHOU Zhi-Yong, ZHAO Song-Hua, ZHANG Wen-Xiang, LIU Gang, ZHAO Zong-Sheng, JIA Bin
. Expression levels of Slc7a11 in the skin of Kazakh sheep with different coat colors[J]. Hereditas(Beijing), 2012
, 34(10)
: 1314
-1319
.
DOI: 10.3724/SP.J.1005.2012.01314
[1] Lamoreux ML, Wakamatsu K, Ito S. Interaction of major coat color gene functions in mice as studied by chemical analysis of eumelanin and pheomelanin. Pigm Cell Res, 2011, 14(1): 23-31.
[2] Sato H, Tamba M, Ishii T, Bannai S. Cloning and expres-sion of a plasma membrane cystine ⁄ glutamate exchange transporter composed of two distinct proteins. J Biol Chem, 1999, 274(17): 11455-11458.
[3] Kim JY, Kanai Y, Chairoungdua A, Cha SH, Matsuo H, Kim DK, Inatomi J, Sawa H, Ida Y, Endou H. Human cystine/glutamate transporter: cDNA cloning and upregulation by oxidative stress in glioma cells. Biochim Biophys Acta, 2001, 1512(2): 335-344.
[4] 苗鲁旭, 孙守荣, 席冬梅, 催嘉, 何奕多, 毛华明, 邓卫东. Slc7a11基因与动物色素的形成. 家畜生态学报, 2009, 30(4): 1-5.
[5] Chintala S, Li W, Lamoreux ML, Ito S, wakamatsu K, Sviderskaya EV, Bennett BC, Park YM, Gahl WA, Huizing M, Spritz RA, Ben S, Novak EK, Tan J, Swank RT. Slc7a11 gene controls production of pheomelanin pigment and proliferation of cultured cells. Proc Nati Acad Sci USA, 2005, 102(31): 10964-10969.
[6] Fontanesi L, Beretti F, Riggio V, Dall’Olio S, Calascibetta D, Russo V, Portolano B. Sequence characterization of the melanocortin 1 receptor (MC1R) gene in sheep with different coat colours and identification of the putative e al-lele at the ovine Extension locus. Small Ruminant Res, 2010, 91(2-3): 200-207.
[7] Norris BJ, Whan VA. A gene duplication affecting expres-sion of the ovine ASIP gene is responsible for white and black sheep. Genome Res, 2008, 18(8): 1282-1293.
[8] Gratten J, Beraldi D, Lowder BV, Mcrae AF, Visscher PM, Pemberton JM, Slate J. Compelling evidence that a single nucleotide substitution in TYRP1 is responsible for coat-colour polymorphism in a free-living population of Soay sheep. Proy Soc B-Bio Sci, 2007, 274(1610): 619-626.
[9] Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2-DDCT method. Methods, 2001, 25(4): 402-408.
[10] PfaffI MW. A new mathematical model for relative quan-tification in real-time RT-PCR. Nucleic Acids Res, 2001, 29(9): 2003-2007.
[11] Dahlin A, Royall J, Hohmann JG, Wang J. Expression pro-filing of the solute carrier gene family in the mouse brain. J Pharmacol Exp Ther, 2009, 329(2): 558-570.
[12] Wagner CA, Lang F, Bröer S. Function and structure of heterodimeric amino acid transporters. Am J Physiol Cell Physiol, 2001, 281(4): C1077-C1093.
[13] Ito S. A chemist's view of melanogenesis. Pigm Cell Res, 2003, 16(3): 230-236.
[14] Wakamatsu K, Ito S. Advanced chemical methods in melanin determination. Pigm Cell Res, 2002, 15(3): 174-183.
[15] He X, Li HT, Zhou ZY, Zhao ZS, Li W. Production of brown/yellow patches in the SLC7A11 transgenic sheep via testicular injection of transgene. J Genet Ge-nome, 2012, 39(6): 281-285.