嗜酸粒细胞__(期刊论文)

MIP4的突变体构建及其对嗜酸粒细胞趋化的抑制

http://www.100md.com 《第四军医大学学报》 2003年第6期

嗜酸粒细胞,,趋化因子；嗜酸粒细胞；基因表达；遗传载体,0引言,1材料和方法,2结果,3讨论,【参考文献】

     (第四军医大学：西京医院呼吸内科，陕西西安 710033，基础部生物化学与分子生物学教研室，科研部生物技术中心)

    Inhibition of eosinophil chemotaxin by mutant of macrophage inflammatory protein 4

    LI ShuJun, QI HaoWen, YE JiangFeng, DI XinYu, JI ShaoPing, HE Peng, ZHANG YingQi, YAO LiBo

    Department of Respiratory Diseases, Xijing Hospital, Department of Biochemistry & Molecular Biology, Faculty of Preclinical Medicine, Center of Biotechnology, Research Administration, Fourth Military Medical University, Xi’an 710033, China

    【Abstract】AIM: To investigate the activity of MIP4 mutant (MetMIP4) in blocking eosinophils chemotaxis in vitro by expressing MetMIP4 in E. coli and purifying it. METHODS: The gene of MetMIP4 was inserted into the expression vector pRSETA and pRSETMetMIP4 fusion protein was expressed by IPTG induction. After E. coli. was lysed by ultrasonic wave, the solubility of the fusion protein was determined by SDSPAGE gel electrophoresis. Protein in supernatant was purified by Ni2+NTA agarose beads. The activity of pRSETMetMIP4 protein in inhibiting eosinophils chemotaxis was determined with 24well chemotaxis plate filter. RESULTS: The pRSETMetMIP4 fusion protein was expressed in E. coli. The solubility of the fusion protein reached 80% and the purity of the fusion protein was 87% by Ni2+NTA agarose beads. Eosinophil was purified from asthmatic blood and Eosinophil purity was 90%. Investigation of biological function of the fusion protein showed that the fusion protein inhibited the eosinophils chemotaxis. CONCLUSION: The pRSETMetMIP4 protein has been correctly expressed in E. coli and the fusion protein is soluble. Study of the biological function of the fusion protein indicates that the fusion protein can inhibit eosinophils chemotaxis in vitro. ......

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