Preparation and identification of recombinant PTD-maxadilan.
- Author:
Le ZENG
1
;
Rongjie YU
;
Mingfang XU
;
Jiansu CHEN
;
Jingjing WANG
;
Juan LI
Author Information
1. Bioengineering Institute of Jinan University, Guangzhou 510632, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Base Sequence;
Blood-Brain Barrier;
metabolism;
Escherichia coli;
genetics;
metabolism;
Genetic Vectors;
genetics;
Insect Proteins;
biosynthesis;
genetics;
pharmacokinetics;
Mice;
Molecular Sequence Data;
Protein Structure, Tertiary;
Recombinant Fusion Proteins;
biosynthesis;
genetics;
pharmacokinetics;
Vasodilator Agents;
metabolism;
pharmacokinetics
- From:
Chinese Journal of Biotechnology
2009;25(11):1739-1745
- CountryChina
- Language:Chinese
-
Abstract:
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.
