Vectors for transforming Mycoplasma hyopneumoniae, transformed M. hyopneumoniae strains, and use thereof
AssigneeHipra Scientific, S.L.U.
InventorsLuis Gonzalez Gonzalez, Jaume Piñol Ribas, Jordi Montane Giralt, Maria Camats Malet, Enrique Querol Murillo, Marta Sitja Arnau
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Abstract
The present invention relates to mutant strains of Mycoplasma hyopneumoniae and to a method for preparing them. It also relates to carrier vectors which are used in said method, to vaccine compositions and to vaccination kits comprising the compositions against porcine enzootic pneumonia and other swine diseases. It also relates to the use of M. hyopneumoniae as a host for expressing recombinant proteins and other DNA sequences of interest.
Claims46
A method for preparing a mutant strain of Mycoplasma hyopneumoniae (Mhyo) comprising transforming a strain of Mhyo with a carrier vector comprising at least one exogenous DNA sequence, wherein the at least one exogenous DNA sequence is under the control of a DNA sequence of a promoter region of Mhyo, wherein the carrier vector is a replicative plasmid vector that comprises:
- 1) a DNA sequence comprising the oriC region of a strain of Mycoplasma sp., which is Mycoplasma hyopneumoniae, and
- 2) an exogenous DNA sequence comprising a marker gene and an additional exogenous DNA sequence, each of which are under the control of a DNA sequence of a promoter region of Mhyo,
- wherein a DNA sequence of a promoter region of Mhyo corresponds to a DNA segment comprising at least 50 base pairs located on the 5′ side of a gene coding for a Mhyo protein,
- wherein a DNA sequence of a promoter region of Mhyo initiates or promotes the transcription of a Mhyo DNA sequence, and
- wherein the exogenous DNA sequence is stably introduced in the cytosol or in the genome of the strain of Mhyo.
A replicative plasmid vector, characterized in that it comprises:
- 1) a DNA sequence comprising the oriC region of a strain of Mycoplasma sp., which is Mycoplasma hyopneumoniae, and
- 2) an exogenous DNA sequence comprising a marker gene, and an additional exogenous DNA sequence, each of which are under the control of a DNA sequence of a promoter region of Mhyo,
- wherein a DNA sequence of a promoter region of Mhyo corresponds to a DNA segment comprising at least 50 base pairs located on the 5′ side of a gene coding for a Mhyo protein, and
- wherein a DNA sequence of a promoter region of Mhyo initiates or promotes the transcription of a Mhyo DNA sequence.
A method for preparing a mutant strain of Mycoplasma hyopneumoniae (Mhyo) comprising transforming a strain of Mhyo with a transposon vector comprising:
- 1) a DNA sequence encoding a transposase,
- 2) an exogenous DNA sequence comprising a marker gene, and
- 3) and an additional exogenous DNA sequence,
- wherein at least one of the exogenous DNA sequences and the DNA sequence encoding the transposase are under the control of a DNA sequence of a promoter region of Mhyo,
- wherein the DNA sequence of the promoter region of Mhyo corresponds to a DNA segment comprising at least a50 base pairs located on the 5′ side of a gene coding for a Mhyo protein, wherein the DNA sequence of the promoter region of Mhyo initiates or promotes the transcription of a Mhyo DNA sequence, and
- wherein the exogenous DNA sequence is stably introduced in the cytosol or in the genome of the strain of Mhyo.
A transposon vector, characterized in that it comprises:
- 1) a DNA sequence encoding a transposase,
- 2) an exogenous DNA sequence comprising a marker gene, and
- 3) an additional exogenous DNA sequence,
- wherein at least one of the exogenous DNA sequences and the DNA sequence encoding the transposase are under the control of a DNA sequence of a promoter region of Mhyo,
- wherein the DNA sequence of the promoter region of Mhyo corresponds to a DNA segment comprising at least 50 base pairs located on the 5′ side of a gene coding for a Mhyo protein, and
- wherein the DNA sequence of the promoter region of Mhyo initiates or promotes the transcription of a Mhyo DNA sequence.
Dependencies are inferred from the claim text.