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Recombinant Toxoplasma fusion protein antigen and preparation process and use thereof

A fusion protein and protein antigen technology, applied in the field of genetic engineering technology and diagnostic reagents, can solve the problem of low antigen specificity

Inactive Publication Date: 2005-03-30
李越希
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Most of the Toxoplasma gondii protein antigens used in my country are cultured Toxoplasma gondii extracts, which contain multiple protein products. The antigen specificity is not strong, and false positives are prone to occur.

Method used

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  • Recombinant Toxoplasma fusion protein antigen and preparation process and use thereof
  • Recombinant Toxoplasma fusion protein antigen and preparation process and use thereof
  • Recombinant Toxoplasma fusion protein antigen and preparation process and use thereof

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Embodiment Construction

[0132] Detailed description of the embodiment of the present invention:

[0133] Fusion expression of Toxoplasma gondii GRA6 protein and P30 protein

[0134] The GRA6 protein gene fragment and the P30 protein gene fragment of Toxoplasma gondii were amplified by PCR method. The two gene fragments were cloned into the NcoI / BamHI and BamHI / EcoRI sites in the same plasmid pET28a(+), so that the two were connected in series, and the translation frame was consistent, and a fusion protein could be expressed. The recombinant plasmid was transformed into Escherichia coli BL21(DE3), and the engineered bacteria that highly expressed the fusion protein was obtained by screening. The expressed fusion protein accounted for about 28% of the total bacterial protein, and existed in two forms of inclusion body and soluble.

[0135] Materials and Methods

[0136] 1. Strains and plasmids: The host strain BL21(DE3) and the expression vector pET28a(+) are products of Novagen, ...

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Abstract

The present invention relates to recombinant toxoplasma fusion protein, its preparation method and application. It utilizes gene engineering technology to prepare a new-type recombinant toxoplasma protein, i.e. the fusion protein formed from 110 amino acids from 43-th amino acid to 152-th amino acid of toxoplasma GRA6 protein N end and 258 amino acids from 56-th amino acid to 313-th amino acid of P30 protein which are series-connected. The above-mentioned 110 amino acids of GRA6 protein N end is positioned at fusion protein N end, and the 258 amino acids of P30 protein is positioned at C end of the fusion protein, and between them 2 amino acids of Gly and Ser are connected, and one methionine (Met) is added at N end of fusion protein, its total length has 371 amino acids.

Description

technical field [0001] The invention utilizes genetic engineering technology to prepare recombinant toxoplasma fusion protein. The two fragments of the fusion protein come from the Toxoplasma gondii GRA6 protein and the P30 protein respectively. The two fragments are linked and expressed by genetic engineering technology to form a new fusion protein. The resulting new fusion protein can be used as an antigen for Toxoplasma gondii antibody or Antigen detection, etc. The invention relates to the fields of genetic engineering technology and diagnostic reagents. Background technique [0002] Toxoplasmosis, also known as toxoplasmosis, is a zoonotic parasitic disease that belongs to the category of parasitic diseases and is distributed worldwide. The pathogen of toxoplasmosis is a coccidia. The whole life cycle development of Toxoplasma needs two hosts. Feline animals, such as domestic cats, are the final hosts, but its selection of intermediate hosts is extremely loose, regard...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K19/00C12N15/62C12N15/63G01N33/569G01N33/577G01N33/68
Inventor 李越希陶开华
Owner 李越希
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