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微管相关蛋白2微管结合区基因的克隆与表达
郭 燕1, 张彦明1, 高 晨2
1.西北农林科技大学 动物科技学院;2.中国疾病预防控制中心 病毒病预防控制所
摘要:
为了研究微管相关蛋白2(MAP2)在促进微管形成、维持微管稳定及促进轴突和树突细胞器转运等方面的作用,试验克隆了MAP2微管结合区基因,构建了原核表达载体pET32a-MAP2并对其进行诱导表达,利用亲和层析法对表达产物进行纯化,研究了表达产物与微管蛋白之间的作用。结果表明,融合表达的人MAP2微管结合区蛋白约43 ku,可与天然的微管蛋白发生分子间的结合作用,为MAP2生理功能的研究奠定了基础。
关键词:  表达纯化  微管蛋白
DOI:
分类号:
基金项目:国家自然科学基金项目(30070038)
Expression and purification of human MAP2 microtubule binding peptide
Abstract:
In order to study the effects of MAP2 on the formation and stabilization of microtubule as well as the potential transportation functions in axon and synapse,the gene encoding microtubule binding region of MAP2 was cloned and expressed.Verified by sequence analysis and enzyme digested,an recombinant vector pET32a-MAP2 was constructed.A 43 ku soluble MAP2-fusion protein was expressed in E.coli BL21 and purified by affinity chromatography.The interaction of the proteins was studied and it was suggested that MAP2 microtubule binding peptide could interact with each other in vitro.It can provid a good basis for further research on the biological functions of MAP2.
Key words:  MAP2  expression and purification  tubulin