Slc7a11基因在不同被毛颜色哈萨克羊皮肤组织中的表达分析

doi:10.3724/SP.J.1005.2012.01314

遗传 ›› 2012, Vol. 34 ›› Issue (10): 1314-1319.doi: 10.3724/SP.J.1005.2012.01314

Slc7a11基因在不同被毛颜色哈萨克羊皮肤组织中的表达分析

李洪涛^1,3, 何新², 周志勇², 赵松华¹, 张文祥¹, 刘钢³, 赵宗胜¹, 贾斌¹

1. 石河子大学动物科技学院, 石河子832003 2. 中国科学院遗传与发育生物学研究所, 北京100101 3. 石河子大学医学院, 石河子832003

收稿日期:2012-06-22 修回日期:2012-08-20 出版日期:2012-10-20 发布日期:2012-10-25
通讯作者: 赵宗胜 E-mail:zhaozongsh@shzu.edu.cn
基金资助:
转基因生物新品种培育重大专项(编号：2009ZX08009-158B)资助

Expression levels of Slc7a11 in the skin of Kazakh sheep with different coat colors

LI Hong-Tao^1,3, HE Xin², ZHOU Zhi-Yong², ZHAO Song-Hua¹, ZHANG Wen-Xiang¹, LIU Gang³, ZHAO Zong-Sheng¹, JIA Bin¹

1. College of Animal Science and Tecnology, Shihezi University, Shihezi 832003, China 2. Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China 3. Shihezi University School of Medicine, Shihezi 832003, China

Received:2012-06-22 Revised:2012-08-20 Online:2012-10-20 Published:2012-10-25

摘要/Abstract

摘要： Slc7a11 基因属于溶质转运家族, 编码胱氨酸/谷氨酸xCT 转运载体, 经证实该基因调控黑色素与伪黑色素的转换。文章利用实时荧光定量PCR技术分析Slc7a11基因在3种不同毛色哈萨克绵羊羔羊皮肤组织中的mRNA转录水平, 构建原核表达载体, 诱导表达融合蛋白, 并对包涵体蛋白进行纯化, 免疫新西兰大白兔制备抗血清, 最后检测不同毛色皮肤中该蛋白的表达水平。结果表明, Slc7a11基因在3种毛色皮肤中的表达水平有显著差异, 棕色被毛皮肤中表达水平最高, 其次是黑色, 在白色中表达最少。利用纯化的融合蛋白制备多克隆抗体, 发现sxCT蛋白在棕色被毛中表达最高, 其次是黑色, 白色最低, 因此, Slc7a11 基因可能与哈萨克绵羊毛色表型有相关性。

关键词: 哈萨克绵羊, Slc7a11, 融合蛋白, 多克隆抗体

Abstract: 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.

Key words: Kazakh sheep, Slc7a11, fusion protein, polyclonal antibody

李洪涛何新周志勇赵松华张文祥刘钢赵宗胜贾斌. Slc7a11基因在不同被毛颜色哈萨克羊皮肤组织中的表达分析[J]. 遗传, 2012, 34(10): 1314-1319.

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, 2012, 34(10): 1314-1319.

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Slc7a11基因在不同被毛颜色哈萨克羊皮肤组织中的表达分析

Expression levels of Slc7a11 in the skin of Kazakh sheep with different coat colors

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