Expression in Spiroplasma citri of an epitope carried on the G fragment of the cytadhesin P1 gene from Mycoplasma pneumoniae.
We have previously described the use of the replicative form (RF) of Spiroplasma citri virus SpV1 as a vector for cloning and expressing foreign genes in S. citri, an organism which reads UGA as a tryptophan codon (C. Stamburski, J. Renaudin, and J.M. Bové, J. Bacteriol. 173:2225-2230, 1991). We now report cloning and expression in S. citri of the G fragment of cytadhesin P1 gene from Mycoplasma pneumoniae. The G fragment was inserted in the SpV1 RF downstream of a synthetic ribosome binding site and introduced into S. citri by electroporation. Northern (RNA) blot analyses showed that in S. citri, the G fragment was transcribed from an SpV1 RF promoter as a 1.2-kb mRNA. The translation product was detected by Western blotting (immunoblotting) with a rabbit antiserum raised against total proteins from M. pneumoniae (strain FH) and was proved to be P1 specific by using monoclonal antibodies specific for the G region of the P1 protein. The apparent molecular mass of the polypeptide (24.5 kDa) indicates that in S. citri, the G fragment was fully translated in spite of the seven UGA codons present in the reading frame.
Marais
A
A
Laboratoire de Biologie Cellulaire et Moléculaire, Institut National de la Recherche Agronomique, Villenave d'Ornon, France.
Bove
J M
JM
Dallo
S F
SF
Baseman
J B
JB
Renaudin
J
J
D018829:Adhesins, Bacterial; D001426:Bacterial Proteins / Q000235:genetics*; D001435:Bacteriophages / Q000235:genetics; D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods*; D000939:Epitopes / Q000235:genetics; D005798:Genes, Bacterial / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics; D008969:Molecular Sequence Data; D009177:Mycoplasma pneumoniae / Q000235:genetics*; D014176:Protein Biosynthesis; D013149:Spiroplasma / Q000235:genetics*; D014158:Transcription, Genetic
D018829:Adhesins, Bacterial; D001426:Bacterial Proteins; D000939:Epitopes; C035471:adhesin, Mycoplasma pneumoniae