Expression of Mycoplasma proteins carrying an affinity tag in M. pneumoniae allows rapid purification and circumvents problems related to the aberrant genetic code.
In Mycoplasma pneumoniae and several other mollicutes, the UGA opal codon specifies tryptophan rather than a translation stop. This often makes it difficult to express Mycoplasma proteins in heterologous hosts. In this work, we demonstrate that mollicute proteins can be fused to an affinity tag and be expressed directly in M. pneumoniae. The protein can then be purified by affinity chromatography and be used for biochemical or any other desired analysis.
Schmidl
Sebastian R
SR
Department of General Microbiology, Institute of Microbiology and Genetics, Georg-August University Göttingen, Grisebachstr. 8, D-37077 Göttingen, Germany.
Hames
Claudine
C
Stülke
Jörg
J
D001426:Bacterial Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D002846:Chromatography, Affinity; D018388:Codon, Terminator / Q000235:genetics; D005815:Genetic Code / Q000235:genetics*; D009174:Mycoplasma / Q000235:genetics*; D009177:Mycoplasma pneumoniae / Q000235:genetics*; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism
D001426:Bacterial Proteins; D018388:Codon, Terminator; D011994:Recombinant Proteins