PMID- 10098689
OWN - NLM
STAT- MEDLINE
DCOM- 19990517
LR  - 20190522
IS  - 1040-6387 (Print)
IS  - 1040-6387 (Linking)
VI  - 11
IP  - 2
DP  - 1999 Mar
TI  - Cloning of Mycoplasma synoviae genes encoding specific antigens and their use as 
      species-specific DNA probes.
PG  - 162-9
AB  - A genomic library of Mycoplasma synoviae (MS) was generated by using 
      bacteriophage lambda gt11 as a cloning and expression vector. Identification of 
      recombinant clones highly specific to MS was achieved by screening the library 
      for expression of MS proteins with polyclonal antiserum that had been preadsorbed 
      with 6 heterologous avian mycoplasma species antigens. Expression of the 
      recombinant clones in Escherichia coli followed by sodium dodecyl 
      sulfate-polyacrylamide gel electrophoresis of the total cell lysates and 
      immunoblot yielded a predominant reactive fusion protein of 165 kD. Two clones 
      (MS2/28 and MS2/12) that yielded inserts of different size were selected. The 2 
      MS DNA inserts were subcloned in a plasmid vector, labeled with digoxigenin, and 
      used as probes for the specific recognition of several MS strains. A high degree 
      of conservation was demonstrated for the MS2/12 and MS2/28 genes in tested MS 
      strains. In addition, neither DNA fragment recognized any other avian mycoplasma 
      species (M. gallisepticum, M. meleagridis, M. gallinarum, M. iners, M. anatis, 
      and M. iowae), thus indicating their high specificity to MS. The sensitivity of 
      the slot blot hybridization method using digoxigenin-labeled MS2/12 and MS2/28 
      probes for direct detection of MS from broth cultures of field isolates was 10(5) 
      colony-forming units/ml. These results demonstrate the effectiveness of adsorbed 
      antisera for the isolation of species-specific mycoplasma DNA and the potential 
      for its use as probes for the specific and direct detection of MS from broth 
      cultures of field isolates.
FAU - Ben Abdelmoumen, B
AU  - Ben Abdelmoumen B
AD  - Departement de Pathologie et Microbiologie, Faculte de Medecine Veterinaire, 
      Universite de Montreal, Saint-Hyacinthe, Quebec, Canada.
FAU - Roy, R S
AU  - Roy RS
FAU - Brousseau, R
AU  - Brousseau R
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - J Vet Diagn Invest
JT  - Journal of veterinary diagnostic investigation : official publication of the 
      American Association of Veterinary Laboratory Diagnosticians, Inc
JID - 9011490
RN  - 0 (Antigens, Bacterial)
RN  - 0 (DNA Probes)
SB  - IM
MH  - Animals
MH  - Antigens, Bacterial/*genetics
MH  - Bacteriophage lambda
MH  - *Cloning, Molecular
MH  - DNA Probes/*genetics
MH  - Escherichia coli/genetics
MH  - Gene Library
MH  - Mycoplasma/*genetics
MH  - Plasmids
MH  - Poultry
MH  - Poultry Diseases/diagnosis/*microbiology
MH  - Species Specificity
EDAT- 1999/03/31 00:00
MHDA- 1999/03/31 00:01
CRDT- 1999/03/31 00:00
PHST- 1999/03/31 00:00 [pubmed]
PHST- 1999/03/31 00:01 [medline]
PHST- 1999/03/31 00:00 [entrez]
AID - 10.1177/104063879901100210 [doi]
PST - ppublish
SO  - J Vet Diagn Invest. 1999 Mar;11(2):162-9. doi: 10.1177/104063879901100210.