Preparation, optimization and characterization of a polyphenylalanine synthetizing system from Agrobacterium tumefaciens.
A very active cell-free translation system was prepared from Agrobacterium tumefaciens, a bacterium broadly used to transfect plant cells to introduce foreign genes and one that produces tumours in plants. Once optimized for Mg2+, NH4+, high speed supernatant S 370, purified ribosomes and time, the system translates polyuridylic acid very efficiently. A. tumefaciens purified ribosomes were inhibited in vitro by several well-known translational inhibitors including some ribosome-inactivating proteins. Treatment of A. tumefaciens purified ribosomes with type 1-RIP crotin 2 lead to the depurination of the 23S rRNA which, upon treatment with acid aniline, released a diagnostic RNA fragment of about 235 nucleotides.
Alegre
C
C
Departamento de Bioquimíca, Biología Molecular y Fisiología, Facultad de Ciencias, Universidad de Valladolid, Spain.
Iglesias
R
R
Ferreras
J M
JM
Carbajales
M L
ML
Girbés
T
T
D016960:Agrobacterium tumefaciens / Q000737:chemistry*; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D002474:Cell-Free System; D009699:N-Glycosyl Hydrolases; D010452:Peptide Biosynthesis; D010455:Peptides; D010940:Plant Proteins / Q000494:pharmacology; D011072:Poly U / Q000235:genetics; D014176:Protein Biosynthesis / Q000187:drug effects*; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology*; D012329:RNA, Bacterial / Q000187:drug effects; D012338:RNA, Ribosomal, 23S / Q000187:drug effects; D054789:Ribosome Inactivating Proteins, Type 1; D054790:Ribosome Inactivating Proteins, Type 2; D012270:Ribosomes / Q000187:drug effects* / Q000378:metabolism; D012276:Ricin / Q000494:pharmacology; D014461:Ultracentrifugation
D000900:Anti-Bacterial Agents; D010455:Peptides; D010940:Plant Proteins; D011500:Protein Synthesis Inhibitors; D012329:RNA, Bacterial; D012338:RNA, Ribosomal, 23S; D054789:Ribosome Inactivating Proteins, Type 1; D054790:Ribosome Inactivating Proteins, Type 2; C079388:melonin protein, Cucumis melo L.; D011072:Poly U; C012001:polyphenylalanine; C012857:crotin II protein, Croton tiglium; D012276:Ricin; D009699:N-Glycosyl Hydrolases