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猪白细胞介素-6基因在大肠杆菌中的表达及其
产物的促淋巴细胞增殖活性
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范俊娟1，陈建1，余兴龙2，张茂林3*，涂长春4
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（1.辽宁农业职业技术学院，辽宁 营口115009；2.湖南农业大学，湖南 长沙410128；3.吉林大学
人兽共患病研究所，吉林 长春130062；4.军事医学科学院 军事兽医研究所，吉林 长春130062）
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摘要：
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为研究猪白细胞介素-6（IL-6）作为免疫佐剂的可行性，用IPTG诱导含有猪IL-6基因的BL21（DE3）LysS工程菌，表达的目的蛋白经超声波破碎，洗涤，8mol/L尿素变性溶解后，用固化金属亲和层析法纯化后用谷胱甘肽再氧化还原法进行复性，并用MTT法对复性的重组蛋白的生物学活性进行分析。结果显示，复性的重组猪IL-6具有良好的刺激淋巴细胞体外增殖的活性。说明重组猪IL-6在作为免疫佐剂方面有一定的应用价值。 |
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关键词：
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猪；IL-6基因；表达；纯化；复性；淋巴细胞增殖活性 |
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中图分类号：Q 511：Q 786 文献标识码：
A 文章编号：1673-4696(2008)06-0531-04 |
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Expression of porcine IL-6 gene in Escherichia coli and lymphocyte
proliferation activity of its expressed product |
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FAN Jun-juan1,CHEN Jian1,YU
Xing-long2,ZHANG Mao-lin3,TU Chang-chun4 |
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(1.Liaoning Agricultural Vocation-technical College,Yingkou 115009,China;2.Hunan Agricultural University,
Changsha 410128,China;3.Institute of Zoonoses,Jilin University,Changchun 130062,China;4.Institute of
Military Veterinary Sciences,The Academy of Military Medical Sciences,Changchun 130062,China) |
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Abstract: To explore the feasibility of porcine interleukin-6 protein(pIL-6) as immunoadjuvant,Escherichia coli BL21(DE3) LysS with the gene encoding pIL-6 was induced by IPTG.The expressed inclusion bodies were solubilized and denatured in the presence of 8mol/L urea，purified by Immoble Metal Affinity Chromatography(IMAC) and renatured with GSH/GSSH.The lymphocyte proliferation activity of the renatured protein pIL-6 was analyzed by using MTT colorimetric analysis.The MTT result showed that the renatured pIL-6 had good stimulating activity for lymphocyte proliferation,indicating that the pIL-6 may have potential as immunoadjuvant. |
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Key words: porcine；IL-6 gene;expression;purification;renaturation;lymphocyte proliferation activity |
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