Toxoplasma wx2 gene deletion strain, construction method and application

A technology of gene deletion and construction method, applied in the field of genetic engineering, can solve problems such as unclear function and mechanism

Pending Publication Date: 2020-06-19
CENT SOUTH UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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  • Toxoplasma wx2 gene deletion strain, construction method and application
  • Toxoplasma wx2 gene deletion strain, construction method and application
  • Toxoplasma wx2 gene deletion strain, construction method and application

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Embodiment Construction

[0054] The present invention will be further described below in conjunction with the accompanying drawings and embodiments. It should be noted that the following description is only for explaining the present invention and not limiting its content.

[0055] The result of the in vitro cell experiment of the present invention is divided into two independent sample t tests, and the time series test is used in the in vivo experiment. Significance is set at 0.05; if P<0.05, it is not significant.

[0056] The present invention relates to the experimental work involving Toxoplasma gondii and the biological environment. Cells are all carried out in the biological environment of the P2 experiment. For liquid containing Toxoplasma gondii or dyes containing Toxoplasma gondii, and animal moving parts, after the experiment is fruitful, the cells should be homogenized in a timely manner.

[0057] 1. Experimental dyes

[0058] 1.1 Experimental animals, insect strains, cell lines, plasmids...

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Abstract

The invention discloses a toxoplasma wx2 gene deletion strain, a construction method and application. The wx2 gene of a toxoplasma RH strain is knocked out through CRISPR/Cas9 technology to successfully obtain the wx2 gene deletion strain with pyrimethamine drug resistance. The specific method is that CRISPR plasmid Psag1-Cas9-U6-sgwx2 and drug-resistant fragment wx2-DHFR are constructed, targetedshearing of the CRISPR plasmid and homologous recombination of the drug-resistant fragment are utilized, the identification of a monoclonal strain and a monoclonal deletion strain is carried out through drug screening, and the wx2 gene deletion strain of the toxoplasma RH strain can be successfully obtained. The toxoplasma wx2 gene deletion strain lays a solid foundation for the study of molecular mechanism of growth of the toxoplasma, and provides a theoretical basis for development of the toxoplasma vaccines and study of the toxoplasma drug target molecules.

Description

technical field [0001] The invention relates to a toxoplasma gene deletion strain, in particular to a toxoplasma wx2 gene deletion strain, a construction method and an application. It belongs to the technical field of genetic engineering. Background technique [0002] The classification of Toxoplasma in the biological kingdom belongs to Apicomplexa, Sporozoa, Eucoccidales, Sarcocystinidae, and Toxoplasma. Toxoplasma parasites are widespread and can infect almost all nucleated cells. Toxoplasma gondii requires the participation of intermediate hosts and terminal hosts to complete the entire life cycle. Toxoplasma gondii is an opportunistic pathogenic protozoan, which is transmitted mainly through fecal-oral, raw or half-cooked meat, and vertical transmission. More than one-third of the world's people are infected with this pathogen. Toxoplasma not only endangers human health, but also is not conducive to animal reproduction and the development of animal husbandry. [0003...

Claims

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Application Information

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IPC IPC(8): C12N1/11C12N15/85C12N15/66C12Q1/04A61K39/002A61P33/02C12R1/90
CPCC07K14/45C12N15/85C12N15/66C12Q1/04A61K39/002A61P33/02G01N2333/45G01N2800/26
Inventor 吴翔马振荣蒋若兰严康管洁舒衡平
Owner CENT SOUTH UNIV
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