Serpin-3 and functions, preparation method and application thereof

A technology of serine protease and inhibitor, applied in the field of biomedicine

Active Publication Date: 2017-03-08
SHENYANG PHARMA UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] At present, there is no research on the structure, preparation, biological function and functional application of Serpin-3 of the Lepidoptera (Lepidoptera) Serpin-3

Method used

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  • Serpin-3 and functions, preparation method and application thereof
  • Serpin-3 and functions, preparation method and application thereof
  • Serpin-3 and functions, preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0100] Isolation and Purification of Natural Serpin-3

[0101] 1. Method-1

[0102] The tussah silkworm body fluid dissolved in anticoagulant buffer solution was centrifuged to remove blood cells, and the tussah silkworm hemolymph without blood cells was used as a sample for purifying serine protease inhibitor-3. The 200ml sample was subjected to ammonium sulfate fractional precipitation, and 35-50% of the precipitated fraction was dissolved and diluted with citric acid-phosphate buffer system, and used as the initial sample loading solution to be loaded on the heparin affinity chromatography column to remove the influence of impurity proteins. The effluent is loaded on the SP-Sepharose cation exchange column, and washed and eluted, and the component containing the target fragment is loaded on the Q-Sepharose anion exchange column for washing and eluted, and then the target component is further processed by the ConA column. Washing and elution to obtain pure Serpin-3 with nat...

Embodiment 2

[0114] Serpin-3 structure analysis and its gene sequence analysis

[0115] The structure of Serpin-3 was analyzed according to the techniques, methods and means of conventional protein chemistry and molecular biology. The specific methods and means are implemented according to the contents listed in the second content of the invention.

[0116] Obtain the complete nucleotide sequence and amino acid sequence of Serpin-3 (as shown in SEQ ID NO: 1).

[0117] The amino acid sequence and nucleotide sequence of the Serpin-3 mature peptide (as shown in SEQ ID NO: 2) were obtained.

Embodiment 3

[0119] Obtaining recombinant Serpin-3 and its analogs and active fragments using prokaryotic expression system

[0120] This example lists and describes the construction strategy and basic method for expressing Serpin-3 and its analogues and active fragment genes of the present invention in a prokaryotic expression system.

[0121] The expression vector, expression host cell and expression strategy of the prokaryotic expression system are the conventional and general expression vector, expression host cell and expression strategy of genetic engineering expression.

[0122] This implementation is to enable those skilled in the art to understand the present invention more comprehensively, but not to limit the scope of the special claims of the present invention in any way.

[0123] For the separation and purification method of the expression product, the method, principle, strategy, etc. of Example 1 were adopted.

[0124] 1. Expression vector construction of Serpin-3

[0125]...

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Abstract

The invention relates to the technical field of bio-medicine and relates to a structure of serpin-3, an obtaining method and new physiological functions of the serpin-3 and application in the field of bio-medicine. The amino acid and nucleotide sequence of complete sequences of the serpin-3 is shown in SEQ ID NO:1. The amino acid and nucleotide sequence of mature peptide of the serpin-3 is shown in SEQ ID NO:2. The invention further provides an analogue or active fragments of the recombinant serpin-3. The natural and recombinant serpin-3 and the analogue and part of fragments of the serpin-3 have an inhibition effect on serine protease, antimicrobial peptide synthesis and a system activating prophenoloxidase in vitro and vivo. The obtaining method of the natural and recombinant serpin-3 and the part of fragments or analogue of the serpin-3 is conventional, simple and high in yield.

Description

[0001] Technical field: [0002] The invention relates to the technical field of biomedicine, and relates to the structure of serine protease inhibitor-3, its obtaining method, its new physiological function and its application in the field of biomedicine. Specifically, the present invention relates to the structure of serine protease inhibitor-3 and its analogs, active fragments, its preparation and production methods and functions, as well as its detection and diagnosis of microorganisms and related molecular patterns, inhibition of insect hemolymph melanin production, inhibition of Applications in the field of biomedicine such as insect synthesis of antimicrobial peptides, preparation of serine protease inhibitor-3 and its analogs or active fragment antibodies and their applications in the field of biomedicine. [0003] Background technique: [0004] The serine protease inhibitor (Serpin) superfamily is the largest superfamily of protease inhibitors, with a wide variety and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/81C12N15/15C12N15/70C12N15/75C12N15/81C12N15/85C07K1/36C07K1/34C07K1/20C07K1/18C07K16/38A61K38/57A61K39/395A61P35/00A61P1/00A61P29/00
CPCA61K38/00A61K2039/505C07K14/811C07K16/38
Inventor 张嵘张景海吴春福
Owner SHENYANG PHARMA UNIVERSITY
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