2 results about "Pyrimethamine" patented technology

The invention provides a toxoplasma gondii gene deletion strain with an immune protection effect and application of the toxoplasma gondii gene deletion strain, and belongs to the technical field of veterinary parasitology, veterinary immunology and parasite molecular biology. According to the toxoplasma gondii gene deletion strain, a gamma-glutamyl hydrolase gene is deleted. The invention also provides a method for constructing the insect strain. The method comprises the following steps: constructing a circular plasmid pSAG1-Cas9-U6-sgGGH; constructing a repair template containing upstream and downstream homologous arms of the gamma-glutamyl hydrolase gene and a DHFR resistance gene; co-transfecting the annular plasmid and the repair template to a toxoplasma gondii ME49 strain; performing drug screening on pyrimethamine to obtain positive monoclone; and confirming that the GGH gene is completely knocked out through PCR (Polymerase Chain Reaction) detection, sequencing analysis and immunofluorescence detection. The toxoplasma gondii gene deletion strain can provide a remarkable protection effect on toxoplasma gondii infection and is high in safety.

The application discloses a Toxoplasma SRS29C and ROP16 double-gene deletion strain and a construction method and application thereof, relates to the technical field of parasite genetic engineering and molecular biology. The Toxoplasma SRS29C and ROP16 double-gene deletion strain is obtained by knocking out SRS29C and ROP16 genes of the ME49 strain through a CRISPR / Cas9 system. The double-gene deletion strain constructed by the application has drug sensitivity change (sensitivity to ethidium bromide), growth defect phenotype, and only carries a single screening marker, and under in-vitro culture conditions, the double-gene deletion strain presents a significant slow growth trend, and the number of worm-containing bubbles is obviously less than that of the wild type and single-gene deletion strain. The double-gene deletion strain substantially influences the proliferation of the worm body, and provides key data support for exploring the molecular basis of the double-gene deletion leading to the increase of the drug sensitivity of the worm body and for analyzing the regulation mechanism of the drug tolerance related path of the Toxoplasma.