Construction of the first shuttle vectors for gene cloning and homologous recombination in Mycoplasma agalactiae.
Mycoplasma agalactiae is a worldwide ruminant pathogen that causes significant economic losses by inflicting contagious agalactia in sheep and goats. The development of efficient control strategies requires a better understanding of the mycoplasma factors that promote successful infection. However, lack of genetic tools has been a major impediment in studying the pathogenic mechanisms of M. agalactiae. This study describes the identification and cloning of the M. agalactiae origin of replication (oriC) in order to construct the first shuttle vectors for targeted gene disruption, gene complementation and expression studies. Additionally, this report provides the first evidence of the occurrence of homologous recombination and the functionality of heterologous tetM determinant in this pathogen.
Chopra-Dewasthaly
Rohini
R
Institute of Bacteriology, Mycology and Hygiene, Department of Pathobiology, University of Veterinary Medicine Vienna, Veterinärplatz 1, A-1210 Vienna, Austria.
Marenda
Marc
M
Rosengarten
Renate
R
Jechlinger
Wolfgang
W
Citti
Christine
C
D001483:Base Sequence; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D018274:Electroporation; D005798:Genes, Bacterial; D005816:Genetic Complementation Test; D005821:Genetic Techniques; D005822:Genetic Vectors; D042822:Genomic Instability; D045644:Mycoplasma agalactiae / Q000235:genetics*; D011995:Recombination, Genetic; D018741:Replication Origin
D004269:DNA, Bacterial