Gene engineering vaccine used for preventing pig cysticercosis and its preparation method

A genetically engineered vaccine, porcine cysticercosis technology, applied in the field of genetic engineering vaccine for preventing porcine cysticercosis and its preparation

Inactive Publication Date: 2005-07-06
中国人民解放军南京军区联勤部军事医学研究所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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  • Gene engineering vaccine used for preventing pig cysticercosis and its preparation method
  • Gene engineering vaccine used for preventing pig cysticercosis and its preparation method

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preparation example Construction

[0040] The preparation method of the genetic engineering vaccine of the present invention takes the hepatitis B virus core protein as a carrier, utilizes molecular biology techniques and methods, and multiple positions between the 1st amino acid to the 183rd amino acid of the hepatitis B virus core protein (HBc) gene Insert the protective antigen gene of cysticercus porcine into the prokaryotic expression plasmid (the prokaryotic expression plasmid can be: pET28a, pET18, pET32, pQE32), transform the host bacteria, and construct engineering bacteria. Induced expression, obtaining a large amount of target protein and purifying it, can obtain the subunit protein vaccine for preventing porcine cysticercosis; or, using the core protein gene of hepatitis B virus as a carrier, using molecular biology techniques and methods, in the core of hepatitis B virus Cysticercus porcine protective antigen genes are inserted at multiple positions between the 1st amino acid to the 183rd amino acid...

Embodiment 1

[0047] As shown in Figure 1, using molecular biology techniques and methods, the epitopes KETc1(n1):APMSTPSATSVRG and KETc12(n2):GNLLLSCLG were inserted between the 78 and 79 amino acid sequences of HBc149, and the epitope GK-1(n3) : GYYYPSDPNTFYAPPYSA was fused to the 149th amino acid to construct the prokaryotic expression plasmid pET28a-Δc-3n. After the correct sequence was proved by SDS-PAGE and sequencing, the engineered bacteria were cultivated on a large scale and expressed, and the subunit protein was obtained after purification Vaccine, this recombinant protein is named as PCCE, and it is quantified by ultraviolet spectrophotometer.

[0048] Mice were immunized by intramuscular injection, supplemented with Freund's incomplete adjuvant, boosted once 3 weeks later, and then boosted again 4 weeks later, a total of three injections. The sera of the mice in the test group and the control group were collected at 1, 3, 5, 7, and 9 weeks after immunization, and the anti-PCCE ...

Embodiment 2

[0051] On the basis of the above example, using molecular biology techniques and methods, the signal peptide gene sequence of human IL-2 was added to the 5' end of the PCCE protein gene sequence, and cloned into the eukaryotic expression plasmid pVAX3.0 to construct a nucleic acid Vaccine pVAX-S-Δc-3n. After the sequencing is correct, a large number of engineered bacteria are cultivated, the plasmid is extracted, purified through a column, and quantified by an ultraviolet spectrophotometer to obtain a nucleic acid vaccine.

[0052] The one-month-old piglets were immunized with nucleic acid vaccine, and boosted once three weeks later. Two weeks after the booster immunization, 20,000 fresh infectious tapeworm eggs were administered to each pig. Three months later, the pigs were killed by bloodletting. The pathological changes of muscle tissue were dissected to calculate the protection rate.

[0053] The experimental results showed that the relative protection rate of nucleic ac...

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Abstract

A gene engineering vaccine for preventing pig cysticercosis and its preparation process are disclosed. The process includes the steps: making the hepatitis B virus core protein as carrier, inserting cysticercus cellulosae protective antigen gene in multiple locations between the first amino acid to No. 183 amino acid hepatitis B core protein, cloning to procaryon expression plasmid or eucaryon expression plasmid, transforming host bacteria, expressing by inducing aim gene and acquiring aim protein or column purified eucaryon expression plasmid DNA therefore, subunit protein vaccine or nucleic acid vaccine for preventing pig cysticercosis are obtained.

Description

technical field [0001] The invention relates to the hepatitis B virus core antigen (HBcAg) protein or gene as a carrier, which carries the protective antigen epitope of Taenia solium, and constructs a subunit protein vaccine and a nucleic acid vaccine. Background technique [0002] Cysticercosis is a zoonotic parasitic disease caused by the infection of pigs or humans by the larvae of Taenia solium - Cysticercus suis. This disease has been found in 27 provinces, municipalities and autonomous regions in my country. Pigs are the main intermediate host, and humans are not only the intermediate host, but also the only final host. The disease is widespread in our country and brings great harm to animal husbandry and human health. Therefore, strengthening the prevention and treatment of porcine cysticercosis is of great significance in promoting the development of pig industry and improving public health. At present, there is no mature method for the prevention and treatment of ...

Claims

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Application Information

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IPC IPC(8): A61K38/16A61K39/00A61K48/00A61P33/14
Inventor 邓小昭刁振宇吴琳周宗安陶开华刘玉王元伦高键王永山郑纪山
Owner 中国人民解放军南京军区联勤部军事医学研究所
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