| 荣昌猪白细胞介素-2受体γ链基因克隆、表达与鉴定 |
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| 引用本文： | 廖晓丹,郭万柱,吴翼,余光勇,蒋清蓉.荣昌猪白细胞介素-2受体γ链基因克隆、表达与鉴定[J].中国兽医学报,2009,29(12). |
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| 作者姓名： | 廖晓丹 郭万柱 吴翼 余光勇 蒋清蓉 |
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| 作者单位： | 四川农业大学,动物生物技术中心,四川,雅安,625014 |
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| 基金项目： | 国家重点基础研究发展规划(973计划)，教育部长江学者和创新团队发展计划 |
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| 摘 要： | 运用RT-PCR技术从由刀豆蛋白(ConA)诱导培养的荣昌猪外周血单核淋巴细胞(PBMC)扩增出猪白细胞介素-2受体γ链基因的完整开放阅读框(ORF),长约1 107 bp,编码由368个氨基酸残基组成的相对分子质量为41 780的蛋白多肽.荣昌猪IL-2Rγ与人、猕猴、牛、犬、鼠在氨基酸水平上的同源性分别为81.6%,80.8%,85.1%,83.2%和68.8%.运用PCR技术从含荣昌猪IL-2Rγ,开放阅读框序列质粒中扩增其成熟蛋白编码基因,共1 020 bp.将其定向克隆于原核表达载体pET-32a(+)后在E.coli BL21中诱导表达,SDS-PAGE结果显示表达的融合蛋白约为52 000,重组蛋白以包涵体的形式表达,表达产物约占茵体总蛋白的37.6%;Western-blotting分析表明,在相对分子质量52 000处有一奈特异性的带;对γ诱导重组茵进行转录检测得到约1 020 bp的特异性片段.
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| 关 键 词： | 猪白细胞介素-2受体γ链基因 克隆 原核表达 |
Cloning,expressiong and characterization of the Rongchang porcine IL-2 receptor γ chain gene |
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| LIAO Xiao-dan,GUO Wan-zhu,WU Yi,YU Guang-yong,JIANG Qing-rong.Cloning,expressiong and characterization of the Rongchang porcine IL-2 receptor γ chain gene[J].Chinese Journal of Veterinary Science,2009,29(12). |
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| Authors: | LIAO Xiao-dan GUO Wan-zhu WU Yi YU Guang-yong JIANG Qing-rong |
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| Abstract: | The complete open reading frame(ORF)of interleukin-2 receptor γ chain gene was amplified from Rongchang porcine peripheral blood lymphocyte stimulated by ConA,which includes 1 107 bp and encoding 41 780 of petide of 368 amino acids.The amino acid sequence showed 81.6%,80.8%,85.1%,83.2% and 68.8% similarities to that of human,macaca,bos taurus,canis familiar,mus musculus chains,respectively.The gene encoding mature IL-2Rγ protein was amplified by PCR from the recombinant plasmid containing IL-2Rγ,which includes 1 020 bp in all.Then it was cloned directionally into prokaryotic expression vector pET-32a(+)and the fusion expression was induced in E coli BL21.SDS-PAGE demonstrated that the fusion protein expressed in form of inclusion body is approximately 52 000,the product of expression accounts 37.6% of the total bacterium protein.Western-blotting analysis indicated that a specific electrophoresis strip was observed by detection after the recombinant bacteria was transcribed. |
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| Keywords: | porcine IL-2 receptor γ chain gene clone prokaryote expression |
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