Abstract
Porcine β defensin 2(pBD2)is a cationic 37-amino acid antimicrobial peptide with disulfide bonds. Synthetic pBD2 had broad antimicrobial activity against pathogenic bacteria, and thus pBD2 could be a good candidate as a bactericidal agent for pigs. This study reported the successful recombinant expression of pBD2 in Escherichia coli and analysis of its antimicrobial activity, its hemolytic activity, salt-tolerance and thermal stability as well. The pBD2 gene, obtained by RT-PCR using the tongue total RNA as a template and cloned into pET30a expression vector, was transformed into E. coli BL21 (DE3) plysS. The recombinant pBD2 was expressed after induction by IPTG and purified by His tag affinity column with 95% purity. The recombinant pBD2 exhibited antimicrobial activity against both Gram-positive S. aureus and Gram-negative E. coli including the multi-resistant E. coli. The minimum inhibitory concentration (MIC) of recombinant pBD2 against tested bacteria was 10 μg/mL, and the recombinant pBD2 could kill 50% E. coli at 14.39 μg/mL and S. aureus at 21.1 μg/mL. In addition, pBD2 showed low hemolytic activity, salt-tolerance and thermal stability, the properties would be important for its application in practice.
Keywords: Antimicrobial activity analysis, Porcine β defensin 2, Recombinant expression.
Protein & Peptide Letters
Title:Molecular Cloning, Expression and Characterization of the Porcine β Defensin 2 in E. coli
Volume: 20 Issue: 6
Author(s): Chun-li Li, Yan-cong Zhao, Xiao-yan Song, Xian-xian Huang and Wei-dong Zhao
Affiliation:
Keywords: Antimicrobial activity analysis, Porcine β defensin 2, Recombinant expression.
Abstract: Porcine β defensin 2(pBD2)is a cationic 37-amino acid antimicrobial peptide with disulfide bonds. Synthetic pBD2 had broad antimicrobial activity against pathogenic bacteria, and thus pBD2 could be a good candidate as a bactericidal agent for pigs. This study reported the successful recombinant expression of pBD2 in Escherichia coli and analysis of its antimicrobial activity, its hemolytic activity, salt-tolerance and thermal stability as well. The pBD2 gene, obtained by RT-PCR using the tongue total RNA as a template and cloned into pET30a expression vector, was transformed into E. coli BL21 (DE3) plysS. The recombinant pBD2 was expressed after induction by IPTG and purified by His tag affinity column with 95% purity. The recombinant pBD2 exhibited antimicrobial activity against both Gram-positive S. aureus and Gram-negative E. coli including the multi-resistant E. coli. The minimum inhibitory concentration (MIC) of recombinant pBD2 against tested bacteria was 10 μg/mL, and the recombinant pBD2 could kill 50% E. coli at 14.39 μg/mL and S. aureus at 21.1 μg/mL. In addition, pBD2 showed low hemolytic activity, salt-tolerance and thermal stability, the properties would be important for its application in practice.
Export Options
About this article
Cite this article as:
Li Chun-li, Zhao Yan-cong, Song Xiao-yan, Huang Xian-xian and Zhao Wei-dong, Molecular Cloning, Expression and Characterization of the Porcine β Defensin 2 in E. coli, Protein & Peptide Letters 2013; 20 (6) . https://dx.doi.org/10.2174/0929866511320060010
DOI https://dx.doi.org/10.2174/0929866511320060010 |
Print ISSN 0929-8665 |
Publisher Name Bentham Science Publisher |
Online ISSN 1875-5305 |
- Author Guidelines
- Graphical Abstracts
- Fabricating and Stating False Information
- Research Misconduct
- Post Publication Discussions and Corrections
- Publishing Ethics and Rectitude
- Increase Visibility of Your Article
- Archiving Policies
- Peer Review Workflow
- Order Your Article Before Print
- Promote Your Article
- Manuscript Transfer Facility
- Editorial Policies
- Allegations from Whistleblowers
Related Articles
-
Cyclooxygenase-2 Inhibition and Gastric Cancer
Current Pharmaceutical Design Recent Updates on Molecular Genetic Engineering Approaches and Applications of Human Therapeutic Proteins
Current Protein & Peptide Science Pharmacological Treatments for Attention-Deficit/Hyperactivity Disorder (ADHD) in Adults
Current Psychiatry Reviews Zinc-Aceclofenac Complex: Synthesis, Hydrolysis Study and Antiinflammatory Studies
Anti-Inflammatory & Anti-Allergy Agents in Medicinal Chemistry Meet Our Editorial Board Member
Anti-Inflammatory & Anti-Allergy Agents in Medicinal Chemistry Editorial [ Inflammatory Bowel Disease and Inflammation-Associated Colon Cancer:Partners in Crime Guest Editor: Silvio Danese ]
Current Drug Targets An Insight into the Hepatoprotective Activity and Structure-activity Relationships of Flavonoids
Mini-Reviews in Medicinal Chemistry Prevalence of Intestinal Parasites among Patients with Chronic Urticaria in Northern Iran
Infectious Disorders - Drug Targets Network Pharmacology and Integrated Molecular Docking Study on the Mechanism of the Therapeutic Effect of Fangfeng Decoction in Osteoarthritis
Current Pharmaceutical Design A Disposable, Highly Sensitive Biosensing System: Determination of Haptoglobin as a Significant Acute Phase Biomarker
Current Analytical Chemistry Developing Antiangiogenic Peptide Drugs for Angiogenesis-Related Diseases
Current Pharmaceutical Design Diet-induced Obesity Differentially Modulates Cardiac Inflammatory Status in the C57 and FVB Mouse Strains
Current Molecular Medicine Mast Cell-IgE-and Mast Cell-Structural Cell Interactions in Allergic Airway Disease
Current Drug Targets - Inflammation & Allergy Impact of the Transfersome Delivered Human Growth Hormone on the Dermal Fibroblast Cells
Current Pharmaceutical Biotechnology Editorial
Current Medicinal Chemistry Induced pluripotent stem cell-derived mesenchymal stem cells: A leap toward personalized therapies.
Current Stem Cell Research & Therapy Angiogenesis-Related Proteins - Their Role in the Pathogenesis and Treatment of Inflammatory Bowel Disease
Current Protein & Peptide Science A Review of Depsipeptide and Other Histone Deacetylase Inhibitors in Clinical Trials
Current Pharmaceutical Design Role of Iodine, Selenium and Other Micronutrients in Thyroid Function and Disorders
Endocrine, Metabolic & Immune Disorders - Drug Targets Corticosteroids in Sepsis: Pathophysiological Rationale and the Selection of Patients
Endocrine, Metabolic & Immune Disorders - Drug Targets