Eimeria tenella eIF3d gene and application thereof

A kind of Eimeria tenella, tender technology, applied in the application, gene therapy, genetic engineering and other directions, can solve the problem of no research report on the eIF3d gene of Eimeria tenella, and achieve the effect of good antigenicity

Inactive Publication Date: 2013-10-23
SHANGHAI VETERINARY RES INST CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, so far there is no research report on Eimeria tenella eIF3d gene

Method used

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  • Eimeria tenella eIF3d gene and application thereof
  • Eimeria tenella eIF3d gene and application thereof
  • Eimeria tenella eIF3d gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Example 1 Cloning and analysis of full-length cDNA of Eimeria tenella eIF3d gene

[0051] The full-length cDNA of the EteIF3d gene was amplified by RACE technology, and the specific steps were as follows:

[0052] 1. Collection and Purification of Eimeria tenella sporozoites

[0053] (1) Add an appropriate amount of PBS buffer solution to the glass homogenizer, add the diluted purified sporulated oocysts to the homogenizer, grind and break the oocysts through the glass homogenizer until the sporangia are completely released, Microscopic examination until no oocysts remained.

[0054] (2) Transfer the sporangia to a 50mL sterilized centrifuge tube, add an appropriate amount of PBS to mix, centrifuge at 3000r / min for 10min, and discard the supernatant. Then washed twice with PBS, and the precipitate was collected.

[0055] (3) Suspend the precipitate with an appropriate amount of PBS buffer solution, transfer it to a 50mL sterilized Erlenmeyer flask, add chicken bile w...

Embodiment 2

[0083] Example 2 Analysis of Expression Differences of EteIF3d Gene in Different Developmental Stages of Eimeria tenella

[0084]The total RNA of four developmental stages of Eimeria tenella (spored oocysts, unsporulated oocysts, sporozoites, and second-generation merozoites) were extracted, and the unsporulated eggs of Eimeria tenella The first strand of cDNA of cysts, sporulated oocysts, sporozoites, and second-generation merozoites was used as a template, and real-time fluorescent quantitative PCR was used to select 18s rRNA as an internal reference to verify the expression of EteIF3d gene in different developmental stages of Eimeria tenella. expression in the body. Table 3 is the real-time fluorescence quantitative PCR amplification primer sequence. The results showed that the EteIF3d gene was highly expressed in the sporozoite and second-generation merozoite stage, and the expression level was highest in the second-generation merozoite stage (see figure 2 ). figure 2...

Embodiment 3

[0087] Example 3 Prokaryotic expression and antigenicity analysis of EteIF3d gene

[0088] 1. Construction of recombinant plasmids for prokaryotic expression

[0089] Primers were designed according to the open reading frame of EteIF3d gene, and the restriction sites of Hind III and Xhol I were respectively added in the upstream and downstream primer sequences. After the target band was cloned into the vector pGEM-T-easy, the plasmid was extracted for double digestion, and then ligated with the expression vector pET-28b(+) double digested with the same restriction endonuclease to construct a recombinant expression plasmid. After identification by PCR and double enzyme digestion, the target band with the expected size was obtained (see image 3 ). It indicated that the prokaryotic expression plasmid containing EteIF3d gene was constructed successfully. image 3 In the middle, 1. pET28b-Ete IF3d recombinant plasmid Xho l I and Hind III double digestion; 2. pET28b empty vector...

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Abstract

The invention discloses an eimeria tenella gene. The eimeria tenella gene is an eimeria tenella elF3d gene which can code an amino acid sequence shown in SEQ ID NO.1. The invention also discloses a protein coded by the eimeria tenella eIF3d gene. Recombinant protein coded by the eimeria tenella eIF3d gene has a good immune protection effect on avian coccidiosis and is applicable to being developed into a new vaccine or new drug which can be used for preventing and treating the avian coccidiosis.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to an Eimeria tenella eukaryotic translation initiation factor 3d gene (eukaryotic translation initiation factor 3d, eIF3d) and its application. Background technique [0002] Chicken coccidiosis is the most prone parasitic disease in large-scale chicken farms, and its pathogen is Eimeria chicken coccidiosis. Chicken coccidiosis can lead to slow growth of poultry and a reduction in feed conversion rate, indirectly causing economic losses in farming, and if it is severe, it will directly cause a large number of chickens to die, thereby causing major economic losses to farmers. At present, there are reports that chicken coccidiosis causes a huge loss of about 500 million pounds to the breeding industry in the world every year. At present, the poultry breeding industry mainly relies on drugs to prevent and control chicken coccidiosis, but the problem of drug resistance of coccid...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/30C12N15/63C12N1/15C12N1/19C12N1/21C12N5/10C07K14/455C07K16/20C07K16/06A61K48/00A61K39/012A61P33/02
Inventor 韩红玉黄兵孔春林姜连连董辉朱顺海赵其平李婷王晔薛璞舒凡帆
Owner SHANGHAI VETERINARY RES INST CHINESE ACAD OF AGRI SCI
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