title: [Construction of novel thioredoxin fusion protein expression system and the production of recombinant Lf-CATH2].
abstract: The objective of this study was to construct an improved thioredoxin fusion protein expression system, and express the cathelicidin-derived peptide, Lf-CATH2. The improved fusion vector Lf-CATH2-pET32α(-TS) was successfully constructed by firstly deleting the thrombin site and S tag from the pET-32α vector, then inserting the Lf-CATH2 plus a thrombin site instead. Afterwards, Lf-CATH2 was expressed in Escherichia coli as fusion protein. After the cleavage by thrombin, Lf-CATH2 was released and subsequently separated using affinity chromatography. The antimicrobial activity of purified Lf-CATH2 was also examined. The improved expression vector significantly increased enzyme cleavage efficiency by 37%, and Lf-CATH2 could be expressed in high yield and maintain the biological activity. This novel thioredoxin fusion protein expression system enables a quick production of high-yield bioactive cationic peptides like cathelicidins.
authors.forename: Yiling
authors.initials: Y
authors.lastname: Lu
authors.forename: Jiuxiang
authors.initials: J
authors.lastname: Gao
authors.forename: Xue
authors.initials: X
authors.lastname: Qiao
authors.forename: Yipeng
authors.initials: Y
authors.lastname: Wang
authors.forename: Haining
authors.initials: H
authors.lastname: Yu
mesh_terms: D054804:Cathelicidins / Q000096:biosynthesis*; D002846:Chromatography, Affinity; D004926:Escherichia coli; D005822:Genetic Vectors; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis*; D013879:Thioredoxins / Q000235:genetics*
chemical_list: D054804:Cathelicidins; D011993:Recombinant Fusion Proteins; D013879:Thioredoxins