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Vector for transforming Mycoplasma hyopneumoniae, transformed Mycoplasma hyopneumoniae strain and use thereof

A technology for Mycoplasma hyopneumoniae and Mycoplasma hyopneumoniae, which is applied in the direction of introducing foreign genetic material, microorganism-based methods, bacterial antigen components, etc. using a carrier, can solve the problems such as the development of Mycoplasma hyopneumoniae and the method for transforming Mycoplasma hyopneumoniae has not been described yet.

Active Publication Date: 2019-04-16
海博莱科学有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although artificial transformation tools or other genetic tools have been developed for some species of M. hyopneumoniae, these methods have not been developed for M. hyopneumoniae, according to the authors
[0024] Thus, methods for transforming M. hyopneumoniae have not been described in the state of the art, although there have been several unsuccessful attempts to perform such transformations

Method used

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  • Vector for transforming Mycoplasma hyopneumoniae, transformed Mycoplasma hyopneumoniae strain and use thereof
  • Vector for transforming Mycoplasma hyopneumoniae, transformed Mycoplasma hyopneumoniae strain and use thereof
  • Vector for transforming Mycoplasma hyopneumoniae, transformed Mycoplasma hyopneumoniae strain and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0329] Embodiment 1.-select Mycoplasma hyopneumoniae bacterial strain

[0330] The wild-type strain of M. hyopneumoniae used was strain 6314 corresponding to the virulent M. hyopneumoniae isolate from Amer-Girona-Spain and from Iowa State University (Iowa State University). ) (Ames-Iowa-USA) strain 232.

Embodiment 2

[0331] Example 2.-Construction of replicative plasmids

Embodiment 21

[0332] Example 2.1.-Construction of replicative plasmid pOG

[0333] To obtain a replicating plasmid in M. hyopneumoniae, the oriC region of M. hyopneumoniae amplified by PCR was introduced into plasmid pBluescript II SK. Oligonucleotide 5OriCNotI(ATG CGC GGC CGC TTA TTT ATCAGA AAC AGT TAG) (SEQ ID NO: 9) and 3OriCXbaI (AGT C GG GCC C AG CTT GCG CAT CAT TGGATG ATG GAT TC) (SEQ ID NO: 10) was used for this amplification and genomic DNA of M. hyopneumoniae strain J was used as template. The resulting 1.96 kb fragment was then digested with restriction enzymes NotI and XbaI. On the other hand, by PCR using the oligonucleotide 5GmORF (ACT G GG ATC C AT GAA TAT AGT TGA AA ATG) (SEQ ID NO: 11) and 3GmApa (GGAT GG GCC C AG CTT GCG CAT CAT TGG) (SEQ ID NO: 12) and use plasmid pIV-T as a template to amplify the gentamicin resistance gene (aac(6')-aph(2")). Then use the corresponding restriction The 1.8 kb PCR product was digested enzymatically. By PCR using the...

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Abstract

The present invention relates to mutant strains of Mycoplasma hyopneumoniae and to a method for preparing said strains. It also relates to vectors used in said method, to vaccine compositions and to vaccine kits which comprise said strains for use against porcine enzootic pneumonia and other porcine diseases. The invention also relates to the use of M. hyopneumoniae as a host for the expression of recombinant proteins and other DNA sequences of interest.

Description

technical field [0001] The present invention belongs to the field of developing methods and tools for the genetic manipulation of Mycoplasma hyopneumoniae with the aim of producing transformed mutant strains of said bacterial species useful as vaccines against swine diseases. Background of the invention [0002] In the veterinary field, diseases caused by microorganisms such as bacteria, viruses or parasites lead to the death of animals or lead to inefficient growth or fattening processes, resulting in serious economic losses. [0003] One method of protecting an animal from future infection involves administering a vaccine to generate an immune response. [0004] Until now, vaccines based on inactivated microorganisms or live attenuated microorganisms have been the main immunological basis for the control or eradication of most infectious diseases. [0005] However, although these types of vaccines give good results, they have problems such as possible reversion of viralit...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/74A61K39/02C12R1/35
Inventor 路易斯·冈萨雷斯冈萨雷斯豪梅·皮诺尔列巴斯霍尔迪·蒙塔内格拉特玛利亚·卡马特斯马莱特恩里克·奎尔罗穆里洛玛尔塔·西塔亚阿尔诺
Owner 海博莱科学有限公司
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