Unlock instant, AI-driven research and patent intelligence for your innovation.
Construction method and application of toxoplasma lactate dehydrogenase gene knockout strain
What is Al technical title?
Al technical title is built by PatSnap Al team. It summarizes the technical point description of the patent document.
A lactate dehydrogenase and gene knockout technology is applied in the field of construction and application of lactate dehydrogenase gene knockout strains of Toxoplasma gondii, and can solve the problem of toxoplasmosis prevention without a good vaccine and the like
Active Publication Date: 2020-04-14
HUAZHONG AGRI UNIV
View PDF8 Cites 0 Cited by
Summary
Abstract
Description
Claims
Application Information
AI Technical Summary
This helps you quickly interpret patents by identifying the three key elements:
the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more
Image
Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
Click on the blue label to locate the original text in one second.
Reading with bidirectional positioning of images and text.
Smart Image
Examples
Experimental program
Comparison scheme
Effect test
Embodiment 1
[0019] Example 1: Construction of Toxoplasma gondii △LDH1△LDH2 strain
[0020] (1) Starting strain ME49
[0021] ME49 is a type II worm strain of the genus Toxoplasma of the family Toxoplasma in the order Coccidia with two genes of lactate dehydrogenase 1 and lactate dehydrogenase 2. The nucleotide sequences of the lactate dehydrogenase 1 and 2 genes are as follows: Shown in SEQ ID NO: 1 and SEQ ID NO: 2.
[0022] (2) Construction of pSAG1-Cas9-TgU6-sgLDH2 plasmid
[0023] ① Use the gRNA online design website (http: / / www.e-crisp.org / E-CRISP / designcrispr.html) to design the target gene targeting site, and design the gRNA primers according to the designed target sequence:
[0096] ② Filter and sterilize with a 0.22um filter.
[0097] 2.2 Toxicity test of △LDH1△LDH2 double knockout strain on mice
[0098] 1) Use HFF cells to culture Toxoplasma gondii tachyzoites in vitro, until 30-50% of the parasites have escaped from the cells; discard the medium in the original culture bottle, and wash away the escaped parasites and residual medium with PBS , add fresh FBS-free dilution.
[0099] 2) Scrape off the cells with a disposable cell scraper, repeatedly blow and beat the suspension 8-10 times with a 5ml syringe, so that the insect vesicles burst and the worms escape, and filter and purify the worms with a sterile 3μm pore-diameter filter membrane; count the cel...
the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More
PUM
Login to View More
Abstract
The invention discloses a construction method and application of a toxoplasma-gondii lactic dehydrogenasegene knock-out strain. According to the method, a toxoplasma-gondii lactate dehydrogenasegene 1 and a toxoplasma-gondii lactate dehydrogenasegene 2 are knocked out, the nucleotide sequence of the lactate dehydrogenase gene 1 is shown in SEQIDNO:2, the nucleotide sequence of lactate dehydrogenase gene 2 is shown in SEQIDNO:2, and the toxoplasma-gondii gene knock-out strain having the nucleotide sequence shown in SEQIDNO:3 and the nucleotide sequence shown in SEQIDNO:4 at the sites of the lactate dehydrogenase gene 1 and the lactate dehydrogenase gene 2. The gene knock-out strain has the advantages of being small in toxicity and little in reproduction in animals and the like. The method can improve the immunization of animals to toxoplasma gondii and has the potential of becoming a genetic engineering vaccine against toxoplasma gondii.
Description
technical field [0001] The invention relates to a method for constructing a lactate dehydrogenase gene knockout strain of toxoplasma gondii, and the use of the lactate dehydrogenase 1 (LDH1) and lactate dehydrogenase 2 (LDH2) double gene deletion strain constructed by the method . Background technique [0002] Toxoplasma gondii is an obligateintracellular parasite belonging to the apicomplexa protozoa. It is widely distributed all over the world. It is a very important zoonotic pathogen and can cause severe zoonotic toxoplasmosis. In South America and some parts of Europe, the infection rate of humans and animals is as high as 80-90%; in China, the infection rate is generally around 10%. Immunocompromised people are at a very high risk of toxoplasma infection, which can be fatal in severe cases. Women infected with Toxoplasma gondii during pregnancy will have adverse effects on fetal development, causing deformities, mental retardation, premature birth and even fetal deat...
Claims
the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More
Application Information
Patent Timeline
Application Date:The date an application was filed.
Publication Date:The date a patent or application was officially published.
First Publication Date:The earliest publication date of a patent with the same application number.
Issue Date:Publication date of the patent grant document.
PCT Entry Date:The Entry date of PCT National Phase.
Estimated Expiry Date:The statutory expiry date of a patent right according to the Patent Law, and it is the longest term of protection that the patent right can achieve without the termination of the patent right due to other reasons(Term extension factor has been taken into account ).
Invalid Date:Actual expiry date is based on effective date or publication date of legal transaction data of invalid patent.
Login to View More 
Login to View More