重组PTD-maxadilan的制备与鉴定  

Preparation and identification of recombinant PTD-maxadilan

作　　者：曾乐[1] 余榕捷[1] 徐明芳[2] 陈建苏[3] 王静静[1] 李娟[1] 

机构地区：[1]暨南大学生物工程研究所,广州510632 [2]暨南大学生命科学技术学院,广州510632 [3]暨南大学医学院眼科,广州510632

出　　处：《生物工程学报》2009年第11期1739-1745,共7页Chinese Journal of Biotechnology

基　　金：广东省自然科学基金项目(No.06300579);教育部科学技术研究重点项目(No.207141);暨南大学青年基金(No.51208018)资助~~

摘　　要：为了构建能够有效进入血脑屏障的新型融合蛋白PTD-maxadilan(PTD-MAX),设计编码融合蛋白PTD-MAX基因,克隆到表达载体pKYB,构建重组表达载体pKYB-PTD-MAX,转化大肠杆菌ER2566中。采用IPTG诱导由PTD-MAX、内含肽和几丁质组成的融合蛋白的表达。利用IMPACT(Intein Mediated Purification with an Affinity Chitin-binding Tag)介导的纯化系统制备目的融合蛋白PTD-MAX。所得的目的蛋白经激光飞行质谱测定分子量,结果与理论值相符。动物实验结果表明融合蛋白PTD-MAX能够有效穿越血脑屏障,重组PTD-MAX具有比天然maxadilan更显著(P<0.05)的抑制小鼠摄食的作用。融合蛋白PTD-MAX的构建和制备为其生物学功能的深入开发奠定了基础。In order to construct a novel fusion protein PTD-maxadilan（PTD-MAX） that can enter the blood-brain barrier（BBB） efficiently,a new gene encoding PTD-MAX was synthesized and cloned into the expression vector pKYB.The recombinant vector pKYB-PTD-MAX was transformed into Escherichia coli ER2566.The expression of fusion protein consisting of PTD-MAX,intein and chitin binding domain was induced by IPTG and the target PTD-MAX protein was purified using Intein Mediated Purification with an Affinity Chitin-binding Tag system.The molecular weight of PTD-MAX determined by the laser flight mass spectrometry was coherent with its theoretical value.The results of the experiment in vivo indicated that the recombinant PTD-MAX can permeate into BBS and inhibitory effects on the food intake were more significantly than maxadilan（P0.05）.The preparation of PTD-MAX lay the foundation for its further application.

关 键 词：PTD-MAX MAXADILAN 内含肽 血脑屏障 

分 类 号：Q78[生物学—分子生物学] S852.659.6[农业科学—基础兽医学]