Characterization of the promoter of rice plastid psaA-psaB-rps14 operon and the DNA-specific binding proteins.
Transcriptional analysis of the promoter of rice psaA-psaB-rps14 operon was performed with Bal 31 deleted mutants in spinach chloroplast extracts. Two functional promoters in this operon (denoted as "-175" and "-129" promoters) were revealed to be utilized by the spinach transcription machinery; however only the "-175" promoter is utilized in vivo. The existence of psaA promoter-specific binding factors in the chloroplast extracts was indicated by competitive gel retardation assay and exonuclease III protection analysis. After ammonium sulfate fractionation of the extracts followed by heparin-agarose column chromatography, the sequence-specific binding factors were characterized to be 66-kDa and 31-kDa polypeptides. Southwestern analysis demonstrated that the two protein factors interact independently with the psaA promoter-containing DNA fragment.
Institute of Biochemistry, National Yang-Ming Medical College, Taipei, Taiwan, Republic of China.
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Chen
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Cheng
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Chen
D001483:Base Sequence; D002736:Chloroplasts / Q000378:metabolism; D004247:DNA / Q000737:chemistry / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000302:isolation & purification / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D015870:Gene Expression; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009876:Operon; D012275:Oryza / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids; D011401:Promoter Regions, Genetic; D015183:Restriction Mapping; D017384:Sequence Deletion; D014158:Transcription, Genetic; D014675:Vegetables / Q000235:genetics
D004268:DNA-Binding Proteins; D004247:DNA