Zeng Le, Yu Rongjie, Xu Mingfang, Chen Jiansu, Wang Jingjing, Li Juan
Bioengineering Institute of Jinan University, Guangzhou 510632, China.
Sheng Wu Gong Cheng Xue Bao. 2009 Nov;25(11):1739-45.
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 (P<0.05). The preparation of PTD-MAX lay the foundation for its further application.
为构建一种能够高效进入血脑屏障（BBB）的新型融合蛋白PTD-麦克斯迪兰（PTD-MAX），合成了一个编码PTD-MAX的新基因，并将其克隆到表达载体pKYB中。将重组载体pKYB-PTD-MAX转化到大肠杆菌ER2566中。用异丙基-β-D-硫代半乳糖苷（IPTG）诱导由PTD-MAX、内含肽和几丁质结合结构域组成的融合蛋白表达，并用内含肽介导的几丁质亲和结合标签系统纯化目标PTD-MAX蛋白。通过激光飞行质谱法测定的PTD-MAX分子量与其理论值相符。体内实验结果表明，重组PTD-MAX能够渗透进入血脑屏障，对食物摄入的抑制作用比麦克斯迪兰更显著（P<0.05）。PTD-MAX的制备为其进一步应用奠定了基础。
