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Design, expression and characterization of recombinant hybrid peptide Attacin-Thanatin in Escherichia coli

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Abstract

Antimicrobial peptides will be attractive and potential candidates as peptide drugs because of their efficient action against microbes and low toxicity to mammal cells. To improve their antibacterial activity, some modifications needs to be made. In this research, the hybrid peptide gene Attacin-Thanatin with 642 bp in length with preferred codons of E. coli was generated using the technology of Gene splicing by overlap extension. The gene was inserted in-frame into E. coli expression plasmid pET-32a (+) and induced to express in E. coli Rosetta. The recombinant protein was partial purified and its biological activity was determined. Analysis of the E. coli Rosetta induced with IPTG revealed that the molecular weight of fusion protein was approximately 41.8 kDa, which perfectly matched the mass calculated from the amino acid sequence. Biological activity detection showed that this peptide effectively inhibited the growth of the test bacteria including E. coli DH5α, E. coli BL21 (DE3), Salmonella choleraesuis and Staphylococcus aureus. Among these bacteria, the Gram-negative E. coli was the most sensitive. Furthermore, there was minor hemolysis activity for porcine red blood cells. So, the results indicated that the hybrid peptide Attacin-Thanatin could be served as a promising candidate for the chemical antibiotics.

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Acknowledgments

The authors thank Mr. X. F. Liu for providing the agarose beads used for protein purification. Project was funded by grants from Program for Changjiang Scholars and Innovative Research Team in University with grant. No. IRTO 555-5, China Ministry of Education, and Feed Biotechnology Project of Sichuan Province of China with grant No.2007Z06-050.

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Authors and Affiliations

  1. Institute of Animal Nutrition, Key Laboratory of Animal Disease-Resistance Nutrition, Ministry of Education, Sichuan Agricultural University, Ya’an, 625014, China

    Li Na Wang, Bing Yu, Jun He & Dai Wen Chen

  2. Cheng Du Product Quality Supervision and Inspection Institute, Sichuan, China

    Li Na Wang

  3. Animal Biotechnology Center, Sichuan Agricultural University, Ya’an, 625014, China

    Guo Quan Han

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  1. Li Na Wang
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  2. Bing Yu
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  3. Guo Quan Han
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  4. Jun He
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  5. Dai Wen Chen
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Correspondence to Dai Wen Chen.

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Wang, L.N., Yu, B., Han, G.Q. et al. Design, expression and characterization of recombinant hybrid peptide Attacin-Thanatin in Escherichia coli . Mol Biol Rep 37, 3495–3501 (2010). https://doi.org/10.1007/s11033-009-9942-3

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  • DOI: https://doi.org/10.1007/s11033-009-9942-3

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