人微管结合蛋白4微管结合区的体外表达

目的：构建表达人微管结合蛋白4微管结合区（Microtubule-binding domain,MTB）的大肠杆菌工程菌。方法：将1.27Kb编码MTB的DNA片段按正确方向亚克隆到原核生物表达载体pET-3α。得到表达质粒JS3，并转化大肠杆菌BL21（DE3）。结果：IPTG诱导下，表达产物的SDS-PAGE分子量大约是40kD，并能被抗MAP4抗体特异地识别。此外，重组MTB蛋白能在体外结合微管。结论：人微管结合蛋白4微管结合区具有微管结合活性。

Expression of Microtubule Binding Domain of Human Microtubule-associated Protein 4

Objective:To construct the engineered E.coli .strain which expresses microtubule-binding domain (MTB) of human microtubule-associated protein 4 (MAP4).Methods:A 1.27Kb DNA fragment encoded MTB was subcloned into expression vector pET-3a.One expression plasmid JS3 was constructed and transferred into BL21(DE3).Results:The molecular weight of expression products was about 40kD on SDS-PAGE.A specific antibody against MAP4 recognized them on Western Blot.In addition,recombinant MTB proteins bound microtubules in vitro.Conclusion:recombinant MTB proteins of human MAP4 retained the MT-binding activity.