鸡AvBD-6全基因的真核表达.docVIP

 The total AvBD-6 gene expressed by Pichia pastoris# Wenyu Sun, Anshan Shan, Qingquan Ma, Na Dong, Yanping Cao, Liang Wang, Zeyun Wang, Changjiang Yan** 5 10 15 20 25 30 35 40 (Northeast Agricultural University) Abstract: Avian β-defenisin 6 (AvBD-6) is a 67 amino acid-long peptide that plays an important role in the innate immunity of chickens and is widely expressed in most tissues. In this study, the codon-optimized AvBD-6 gene was synthesized and cloned into the eukaryotic expression vector pPICZαA and expressed in Pichia pastoris. Expression levels of the recombinant AvBD-6 were tested by Tricine-SDS, and the product concentration reached 213.0 mg/l after optimization of the expression conditions. Recombinant AvBD-6 exhibited antimicrobial activity against bacteria with the minimal inhibition concentration (MIC) from 5.8 to 46.6 μg/ml and showed no hemolytic activity. Key words: antimicrobial peptides; AvBD-6; total gene sequence; expression; Pichia pastoris 0 Introduction Antibiotic abuse and resistance pose a continuing threat to public health worldwide. Abuse of antibiotics may cause a variety of problems, such as drug resistance and antibiotic residues. Therefore, a new generation of antimicrobials must be developed to meet the global challenge of antibiotic resistance. In response to this problem, antimicrobial peptides (AMPs) have been explored as potential antibiotic alternatives [1]. AMPs were originally identified by their vital role in protecting the body-environment interface from infection. AMPs are an important component of the natural defense system and can act as antibacterial, antifungal, antiviral and even anticancer drugs [2]. Defensins are usually cationic peptides with low molecule weights ranging from approximately 1 to 5 kDa. Defensins are cysteine-rich AMPs, which are broadly divided into five groups: plant, invertebrate, α-, β-, and θ-defensins, with the last three groups being predominantly found in mammalian species [3]. Avian β-