Homotype tandem polymer of BmSPI38 as well as construction method and application of homotype tandem polymer

A construction method and multimer technology, applied in peptide preparation methods, microbial-based methods, chemical instruments and methods, etc., can solve the problems of low multimer activity, poor uniformity, no inhibitory activity, etc., and achieve the expression form Uniform, strong activity, uniform expression effect

Active Publication Date: 2021-12-14
SHAANXI SCI TECH UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, studies have found that the monomeric form of BmSPI38 has no inhibitory activity, and it has been found that although the multimeric form of BmSPI38 has inhibitory activity against subtilisin and proteinase K, the effect of multimerization on the activity of protease inhibitors is not yet fully understood.
And found that the BmSPI38 multimer in the prior art has low activity and poor uniformity

Method used

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  • Homotype tandem polymer of BmSPI38 as well as construction method and application of homotype tandem polymer
  • Homotype tandem polymer of BmSPI38 as well as construction method and application of homotype tandem polymer
  • Homotype tandem polymer of BmSPI38 as well as construction method and application of homotype tandem polymer

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Experimental program
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Effect test

Embodiment 1

[0054] 1. Preparation of experimental materials and reagents

[0055] 1. Experimental materials

[0056] Host bacteria Escherichia coli Trans1-T1 competent cells were purchased from Beijing Quanshijin Biotechnology Co., Ltd., BL21 (DE3) competent cells were purchased from Sangon Bioengineering (Shanghai) Co., Ltd., and Origami 2 (DE3) strains were preserved by our laboratory , the p28 carrier was preserved by the Institute of Vitamin D Physiology and Application of Shaanxi University of Technology.

[0057] 2 experimental reagents

[0058] (1) Reagents related to molecular cloning

[0059] Antibiotic stock solutions and their working concentrations:

[0060] Kanamycin sulfate, stock solution concentration is 50mg / mL (ddH 2 O), the working concentration is 50μg / mL (ddH 2 O), storage condition -20°C;

[0061] Streptomycin, stock solution concentration is 10mg / mL (ddH 2 O), the working concentration is 10 μg / mL (ddH 2 O), storage condition -20°C;

[0062] Tetracycline hy...

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Abstract

The invention relates to the technical field of polymer construction, and particularly discloses a homotype tandem polymer of BmSPI38 as well as a construction method and application of the homotype tandem polymer. Tandem polymer primers BmSPI38-Nde I-BamH I-F, BmSPI38-Not I-R, BmSPI38-Bgl II-R and BmSPI38-L-Bgl II-R are designed and synthesized, BmSPI38-p28 is used as a template for amplification to obtain a target gene segment, a BmSPI38 homotype tandem polymer carrier is designed and constructed by using an isocaudarner method, and carrying out transformation, inducible expression and purification to obtain the BmSPI38 homotype tandem multimer. The obtained BmSPI38 homotype tandem polymer can successfully inhibit the activity of subtilisin and protease K, the activity is higher, and the expression form is more uniform.

Description

technical field [0001] The invention relates to the technical field of multimer construction, in particular to a homotype tandem multimer of BmSPI38 and its construction method and application. Background technique [0002] Bombyx mori (also known as silkworm) is a silk insect belonging to the family Lepidoptera and has very high economic value. Studies have found that the immune process in insects lacks the participation of lymphocytes and immunoglobulins, and pathogenic microorganisms will secrete a large number of extracellular proteases in the process of invading their hosts. the intrusion process. In order to resist the invasion of pathogenic microorganisms, host organisms generally express high levels of protease inhibitors to resist the extracellular proteases secreted by pathogenic microorganisms. Therefore serine protease inhibitors play a very important role in the immune system of insects. [0003] A total of 80 serine protease inhibitors (serine protease inhib...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/70C07K19/00C07K1/22A61K38/57A61P31/10C12R1/19
CPCC12N15/70C07K14/811A61P31/10C07K2319/00A61K38/00
Inventor 李游山杨玺
Owner SHAANXI SCI TECH UNIV
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