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Leptospira interrogans DNA (Deoxyribose Nucleic Acid) vaccine as well as construction method and application thereof

A leptospira and DNA vaccine technology, which is applied in the field of medicine and biology, can solve the problems such as no reports of Leptospira question mark DNA vaccine, and achieves the effects of stable physical and chemical properties, convenient storage and transportation, and simple production process.

Inactive Publication Date: 2011-03-30
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] There are currently no reports of a Leptospira interrogans DNA vaccine

Method used

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  • Leptospira interrogans DNA (Deoxyribose Nucleic Acid) vaccine as well as construction method and application thereof
  • Leptospira interrogans DNA (Deoxyribose Nucleic Acid) vaccine as well as construction method and application thereof
  • Leptospira interrogans DNA (Deoxyribose Nucleic Acid) vaccine as well as construction method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Embodiment 1: The structure, construction and preparation of the DNA vaccine pVAX-LB018 of the present invention:

[0044] The structure of the DNA vaccine of the present invention is a recombinant plasmid DNA molecule (purchased from Invitrogen Company) with the eukaryotic expression vector—pVAX as the backbone, and the general structure is as follows: figure 1 shown. The total length is about 4.0kb (kilobase pairs), and besides containing the pMB1 ori plasmid replication initiation site, kanamycin resistance gene Kanamycin and other elements, it also contains a complete eukaryotic transcription and translation unit. The eukaryotic transcription-translation unit contains elements necessary for eukaryotic transcription and translation: eukaryotic cytomegalovirus promoter P CMV , the corresponding coding gene LB018 and the polyadenylic acid BGH pA at the 3' end. Generally speaking, the vaccine is a recombinant plasmid DNA that can express LB018 in eukaryotic cells and...

Embodiment 2

[0054] Example 2: Functional testing of the DNA vaccine pVAX-LB018 of the present invention:

[0055] 1, obtain the DNA vaccine of the present invention of high purity:

[0056] Take out the frozen engineered strain——DH5α(pVAX-LB018) from the -80°C refrigerator, and after thawing at room temperature, use an inoculation loop to pick up a small amount of bacterial solution and inoculate it on a 1.5% LB agar plate (containing kanamycin). 50ug / ml), after culturing overnight at 37°C, take a single clone and inoculate it in 50ml LB liquid medium (containing 1% tryptone, 0.5% yeast extract and 1% sodium chloride, kanamycin 50ug / ml, pH7 .0), cultured with shaking at 37°C for 40 hours, and used as seed solution. The seed solution was inoculated in 1 liter of SOC liquid medium (on the basis of the above-mentioned components, additionally containing 50 μg / ml kanamycin) at a ratio of 1:20, and cultured with vigorous shaking at 37°C for 25 hours; To a final concentration of 170 μg / ml, cu...

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Abstract

The invention relates to the technical field of medical biology, in particular to a leptospira interrogans DNA (Deoxyribose Nucleic Acid) vaccine as well as a construction method and application thereof. The leptospira interrogans can rapidly invade the inside of human and various animals to cause infection via the skin or the mucous membranes, and result in leptospirosis with the main symptoms of fever, jaundice and bleeding after humans are infected, the leptospirosis is the common zoonosis with wide world distribution and seriously threat the health of the humans and the production of animal husbandry, and China is one of the main countries with the epidemic leptospirosis. The invention provides the leptospira interrogans DNA vaccine, which comprises a eukaryon expression vector and leptospira interrogans LB018 genes, wherein the nucleotide sequence of the LB018 genes is shown as SEQ ID NO: 1. The leptospira interrogans DNA vaccine can effectively induce the immune response reaction of a piggy, which shows that the leptospira interrogans DNA vaccine has a certain immune protection action on the leptospira interrogans of the animals, and the leptospira interrogans DNA vaccine has the advantages of simple production technology, low cost and stable physical and chemical properties, and is convenient for storage and transportation.

Description

technical field [0001] The invention relates to the field of medical biotechnology, in particular to a Leptospira interrogans DNA vaccine and its construction method and application. Background technique [0002] Leptospira interrogans (Letospira interrogans for short) can quickly invade humans and various animals through the skin or mucous membranes to cause infection. Humans can cause leptospirosis with fever, jaundice, and bleeding as the main symptoms. Symptoms are mild or asymptomatic. Due to the large number of natural hosts of Leptospira interrogans and the rapid spread through polluted water sources, leptospirosis is a common zoonotic disease that is widely distributed in the world and seriously threatens human health and animal husbandry production. One of the main endemic countries. [0003] Leptospira vaccine is an important means to prevent leptospirosis. Although the currently used inactivated leptospira vaccine and the trial outer membrane vaccine have a good...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61K39/02C12N15/31C12N15/79A61P31/04
CPCY02A50/30
Inventor 孙树汉刘音郭瀛军丁飞翔单石
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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