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Antibacterial peptide of bactrocera dorsalis hendel and derivatives and application of antibacterial peptide

A technology of Bactrocera dorsalis and antimicrobial peptides, applied in the fields of Bacteralis dorsalis antimicrobial peptides and their derivatives and applications

Inactive Publication Date: 2015-02-18
GUANGZHOU GLAM BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Bactrocerin-2, which is a Diptera insect, should theoretically have an antimicrobial peptide gene in its genome. Dang Xiangli et al. , there are few reports on related research

Method used

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  • Antibacterial peptide of bactrocera dorsalis hendel and derivatives and application of antibacterial peptide
  • Antibacterial peptide of bactrocera dorsalis hendel and derivatives and application of antibacterial peptide
  • Antibacterial peptide of bactrocera dorsalis hendel and derivatives and application of antibacterial peptide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Construction of cDNA library and EST sequence analysis of Bactrocera dorsalis larvae induced by mixed bacteria

[0032] Staphylococcus aureus and Escherichia coli K 12 D. 31 The larvae of Bactrocera dorsalis were needled with mixed bacteria, and after 24 hours of induction, the total RNA of Bactrocera dorsalis larvae was extracted according to the instruction manual of the invitrogen TRIzol Reagent RNA extraction kit. It was observed that the characteristic bands of 18S and 28S were neat and clear, and the brightness ratio was close to 1:2, while measuring OD by UV spectrophotometer 260 / OD 280 =1.82, indicating that the quality of total RNA is better. The library was constructed using the Smart cDNA Library Construction Kit kit from Clontech Company, and experiments were carried out using the primers, vectors, and restriction endonucleases attached to the kit itself. Get the total RNA of 1 μg Bacteralis dorsalis larvae, with SMART III oligonucleotide frag...

Embodiment 2

[0035] Example 2 Recombinant Expression of Bacteralis dorsalis Antimicrobial Peptide BH-AMP003

[0036] In this experiment, the histidine-deficient Pichia pastoris GS115 was used as the host strain, and the intracellular expression vector pPIC3.5K was used to construct the recombinant expression system. Select the Sac I endonuclease to linearize the recombinant vector pPIC3.5K, and then pass the BH-AMP003 gene (shown in SEQ ID NO: 3) with the corresponding restriction site through molecular cloning T4 DNA ligase was ligated with the linearized vector, and then transformed into Escherichia coli DH5α, and positive transformants were screened using ampicillin-resistant LB plates. Pick a single colony for inoculation and culture, and extract the plasmid for enzyme digestion verification and sequencing. For the detailed method of the whole experiment, please refer to "Molecular Cloning Laboratory Manual" edited by Sam brook J. The recombinant vector was further transformed into P...

Embodiment 3

[0063] Example 3 Mouse Acute Toxicity Test of Bacteralis dorsalis Antimicrobial Peptide BH-AMP003 Expressed by Recombinant Yeast

[0064] The recombinant yeast expressing the Bacteralis dorsalis antimicrobial peptide BH-AMP003 prepared above was fermented, and the fermented liquid was spray-dried to obtain a dry bacterial liquid powder of the Bacteralis dorsalis antibacterial peptide BH-AMP003, which was used in subsequent experiments.

[0065] Select 40 healthy NIH mice, male or female, weighing about 30 g, and randomly divide them into two groups, namely, the blank control group and the BH-AMP003 bacteria solution dry powder group, with 20 mice in each group. Before the experiment, the weight of the mice was recorded once. At the beginning of the experiment, the mice in each group were given the test sample at a time with a maximum administration volume of 0.8ml. After the administration, the mice were continuously observed for 7 days to observe whether the mice appeared ment...

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Abstract

The invention discloses antibacterial peptide of bactrocera dorsalis hendel and derivatives and application of the antibacterial peptide. An amino acid sequence of antibacterial peptide is as shown in SEQ ID NO:1; the derivatives of antibacterial peptide are antibacterial active derivatives obtained through modification or antibacterial derivatives formed through substitution, deletion or addition of one or more amino acid residues; obtained antibacterial peptide BH-AMP003 has bioactivity and a wide antibacterial effect, so that antibacterial peptide BH-AMP003 can be applied to preparation of antibacterial products including feed additives, animal drugs, preservatives and medical products such as antibacterial drugs.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a Bactrocera dorsalis antibacterial peptide and its derivatives and applications. Background technique [0002] Insects are one of the very large biological populations. It is estimated that there are about 8 million species of insects in the world, accounting for more than 95% of the total number of animals. There are traces of insects, and these special environments have shown that insects themselves have a strong ability to adapt to survival and defend against the outside world. The defense system of insects is different from that of vertebrates. They do not have a complete B lymphatic system and T lymphatic system. There is no perfect immunoglobulin and complement. A large number of research results show that when insects are infected with pathogenic bacteria or may cause pathogenic bacteria infection, insects themselves can quickly synthesize a large number of antiba...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/435A23K1/16
CPCA23K20/142C07K14/43577
Inventor 李莉李本涛
Owner GUANGZHOU GLAM BIOTECH
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