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Bufo gargarizans lysozyme

A technology of Bufo bufo and lysozyme, applied in the field of biomedicine

Inactive Publication Date: 2017-04-26
ZHEJIANG FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

There are also some peptides that have certain side effects on the body, such as the Maximin H antimicrobial peptide of the Chinese Bombina maxima, the Brevinin-1 antimicrobial peptide of the Japanese marsh frog, and the Bombinin H antimicrobial peptide of the Oriental Bombintororientalis, all of which have strong hemolytic activity.

Method used

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  • Bufo gargarizans lysozyme
  • Bufo gargarizans lysozyme
  • Bufo gargarizans lysozyme

Examples

Experimental program
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Embodiment

[0021] TRIzol reagent method (Invitrogen TM , USA) to extract the total RNA of the glandular tissue, and use the ReverTra AceqPCR RT Kit (TOYOBO, Japan) reverse transcription kit to reverse transcribe the total RNA into cDNA.

[0022] Using primers 5'-CCGGAATTCAAGTATGAAAAATGCGAACTTGCTA-3' and 5'-CCCTTCGAATTAAAGTTTGCATCCTTGAGTCCA-3' with added restriction sites as primers, the cDNA synthesized in the above steps was used as a template to amplify the coding sequence fragment of the lysozyme functional domain of B. sinensis.

[0023] The cloned fragment was purified with a DNA purification kit, and the purified DNA fragment and pET-28 empty plasmid were both digested with EcoR I and Hind III, and the reaction condition was 37°C for 8 hours. After the reaction, the digested product was purified again with a DNA purification kit, and the purified DNA fragment and the pET-28 empty plasmid fragment were ligated with T4 DNA ligase.

[0024] The ligated recombinant plasmid was transfo...

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Abstract

The invention discloses a Bufo gargarizans lysozyme. The Bufo gargarizans lysozyme has a nucleotide sequence shown in SEQ ID NO: 1. The lysozyme recombinant expression product has an inhibiting effect on growth of two kinds of gram positive bacteria, two kinds of gram negative bacteria and two kinds of fungi.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a Bufo bufo lysozyme. Background technique [0002] The ecological and environmental disasters caused by the abuse of antibiotics in today's world and their impact on human health have attracted more and more attention from the world. Finding alternatives to antibiotics with better activity and higher specificity has become a top priority. Therefore, polypeptide substances with antibacterial activity-antimicrobial peptides (AMPs) have become a new research hotspot. Among them, amphibian-derived AMPs represented by Maganin Especially eye-catching. The research on skin secretions of amphibians mostly focuses on the identification and function of peptides. For example, the antimicrobial peptide Magainin of Xenopys laevis and the antimicrobial peptide Maximin of Bombina maxima can kill tumor cells at very low concentrations, and they can selectively act on tumor cells while affecting nor...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/56C12N9/36C12N15/70A61P31/04A61P31/10
CPCC12N9/2462C12N15/70C12Y302/01017
Inventor 高飞钱永常
Owner ZHEJIANG FORESTRY UNIVERSITY
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