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人可溶性TRAIL蛋白表达及纯化条件优化
引用本文:赵金金,徐志辉,胡磊,罗文雅,胡威,尤红娟,李向阳,颜超,郑葵阳,汤仁仙.人可溶性TRAIL蛋白表达及纯化条件优化[J].徐州医学院学报,2014(5):302-305.
作者姓名:赵金金  徐志辉  胡磊  罗文雅  胡威  尤红娟  李向阳  颜超  郑葵阳  汤仁仙
作者单位:徐州医学院病原生物学与免疫学教研室,江苏徐州221004
基金项目:基金项目:江苏省普通高校研究生科研创新计划(CXZZ12-0986);徐州市科技计划项目(XZZD1331)
摘    要:目的:优化人可溶性TRAIL蛋白生产菌发酵及纯化条件,从而提高TRAIL的产量及活性。方法①探索6种不同培养液、培养液pH值、异丙基硫代半乳糖苷( IPTG)诱导浓度及时间,选择最佳发酵条件;②联合镍离子亲和层析及超滤法纯化人可溶性TRAIL蛋白。结果①人可溶性TRAIL的最佳发酵条件为pH7.4的YPD培养液,所用IPTG的最佳浓度为0.2 mmol/L,最佳诱导时间为6 h;②经镍离子亲和层析和超滤联合作用可得纯度极高的蛋白。结论成功完成了人可溶性TRAIL发酵及纯化条件的优化,为今后大量提取TRAIL蛋白奠定了基础。

关 键 词:TRAIL  原核表达  纯化  优化

The optimization of expression and purification of human soluble TRAIL protein
ZHAO Jinjin,XU Zhihui,HU Lei,LUO Wenya,HU Wei,YOU Hongjuan,LI Xiangyang,YAN Chao,ZHENG Kuiyang,TANG Renxian.The optimization of expression and purification of human soluble TRAIL protein[J].Acta Academiae Medicinae Xuzhou,2014(5):302-305.
Authors:ZHAO Jinjin  XU Zhihui  HU Lei  LUO Wenya  HU Wei  YOU Hongjuan  LI Xiangyang  YAN Chao  ZHENG Kuiyang  TANG Renxian
Institution:( Department of Microbiology and Immunology, Xuzhou Medical College, Xuzhou, Jiangsu 221004, China)
Abstract:Objective To optimize the production and purification conditions of human soluble TNF -related apop-tosis inducing ligand ( TRAIL) , thereby increasing the productivity and activity of TRAIL .Methods ①6 different me-dium, pH, the induced concentration and time of isopropyl β-D-1-thiogalactopyranoside ( IPTG) were investigated and the optimum fermentation conditions were selected .②Better human soluble TRAIL protein was obtained through the combination of nickel ion affinity chromatography and ultrafiltration .Results ①The optimum fermentation conditions for human soluble TRAIL was YPD medium at pH 7.4.The optimum concentration of IPTG used was 0.2 mmol/L, the best time was 6 hours.②Through nickel ion affinity chromatography and ultrafiltration combination , higher effects and more purity protein could be obtained .Conclusion The fermentation conditions for human soluble TRAIL are successfully op-timized, which lays foundation for future extraction of large amount of TRAIL protein .
Keywords:TNF -related apoptosis inducing ligand  prokaryotic expression  purification  optimization
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