[Preparation and identification of recombinant PTD-maxadilan].
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.
Zeng
Le
L
Bioengineering Institute of Jinan University, Guangzhou 510632, China.
Yu
Rongjie
R
Xu
Mingfang
M
Chen
Jiansu
J
Wang
Jingjing
J
Li
Juan
J
D000818:Animals; D001483:Base Sequence; D001812:Blood-Brain Barrier / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics; D019476:Insect Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000493:pharmacokinetics*; D051379:Mice; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000493:pharmacokinetics*; D014665:Vasodilator Agents / Q000378:metabolism / Q000493:pharmacokinetics
D019476:Insect Proteins; D011993:Recombinant Fusion Proteins; D014665:Vasodilator Agents; C069147:maxadilan protein, insect