| 麦洼牦牛 Hoxa10基因克隆、原核表达及组织表达谱 |
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| 引用本文： | 李 宇,熊显荣,兰道亮,宋 虎,李 键.麦洼牦牛 Hoxa10基因克隆、原核表达及组织表达谱[J].西北农业学报,2013,22(7):7-11. |
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| 作者姓名： | 李 宇 熊显荣 兰道亮 宋 虎 李 键 |
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| 作者单位： | （西南民族大学 生命科学与技术学院，成都 610041） |
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| 基金项目： | 国家科技支撑计划（2012BAD13B06）。 |
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| 摘 要： | 从麦洼牦牛肾脏组织提取总RNA，以已公布牛 Hoxa10编码区序列设计引物，RT PCR法克隆麦洼牦牛 Hoxa10基因；构建原核表达载体pET 32a(+) Hoxa10，并在大肠杆菌表达系统中表达，SDS PAGE检测融合蛋白；采用RT PCR检测该基因在麦洼牦牛各组织中的表达分布。结果表明：从麦洼牦牛肾脏中克隆 Hoxa10基因的CDs区，大小为285 bp，编码94个氨基酸残基，与牛、猪和绵羊的 Hoxa10同源性高；经IPTG诱导，pET 32a(+) Hoxa10在BL21中可表达1个大小约为28 ku的特异蛋白；在牦牛各组织中，肾脏和肺脏中表达较高，但在心脏表达量极低或不表达。
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| 关 键 词： | Hox基因 表达谱 克隆 原核表达 麦洼牦牛 |
Cloning, Prokaryotic Expression and Tissue Expression Pattern of Hoxa10 in Maiwa Yak |
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| LI Yu,XIONG Xianrong,LAN Daoliang,SONG Hu and LI Jian.Cloning, Prokaryotic Expression and Tissue Expression Pattern of Hoxa10 in Maiwa Yak[J].Acta Agriculturae Boreali-occidentalis Sinica,2013,22(7):7-11. |
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| Authors: | LI Yu XIONG Xianrong LAN Daoliang SONG Hu and LI Jian |
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| Abstract: | Hox gene is a class cluster of genes that specifically regulate biological body and control bone formation in mammals. Total RNA was isolated from kidney of Maiwa Yak (Bos grunniens), designed primers by the coding region of bovine Hoxa10, and cloned the Hoxa10 of Maiwa Yak by RT PCR. The CDs was subcloned into the expression plasmid pET 32a(+) and constructed expression plasmid pET 32a (+) Hoxa10, induced expression in BL21 and detected by SDS PAGE. Moreover, tissue expression patterns were tested by RT PCR. Results showed that the CDs of Hoxa10 was cloned from kidney of Maiwa Yak, about 285 bp and encoding a peptide of 94 amino acid residues, which showed high homology with bovine, pig and sheep. SDS PAGE showed that after IPTG inducing, BL21 containing pET 32a(+) Hoxa10 can express a specific protein of approximately 28 ku;the expression of Hoxa10 in lung and kidney were significant higher than other tissue, however, little or no expression was detected in heart. |
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| Keywords: | Hox gene Expression pattern Cloning Prokaryotic expression Maiwa yak |
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