PMID:1995430
| Citation |
Coppola, G, Huang, F, Riley, J, Cox, JL, Hantzopoulos, P, Zhou, LB and Calhoun, DH (1991) Sequence and transcriptional activity of the Escherichia coli K-12 chromosome region between rrnC and ilvGMEDA. Gene 97:21-7 |
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| Abstract |
We previously identified a protein related to the expression of the ilvGMEDA cluster of Escherichia coli K-12. It was observed that this ilv-related protein was produced at higher levels in UV irradiated cells infected with lambda dilvGMEDA phage with specific ilvG mutations (ValR), compared to phage carrying the wild-type(ValS) ilvG allele. The gene encoding this protein was further localized to a region between rrnC and ilvGMEDA by analyzing restriction fragment subsets in maxicells. We have now determined the nucleotide (nt) sequence of the 3.5-kb segment between rrnC and ilvGMEDA, and two open reading frames (ORFs) are present in the region expected to contain the ilv-related gene. These ORFs predicts Mrs of 18,751 (ORFI) and 20,085 (ORFII) Da, and both ORFs have a strong probability to encode proteins based on codon frequency analysis. Maxicell analysis revealed that a 1319-bp HindIII-SmaI fragment containing ORFI encodes the ilv-related peptide. We deleted a ClaI fragment that removed a portion of ORFI encoding the C-terminal region of the peptide, and maxicell analysis revealed a decrease in the size of the protein produced in accord with the prediction. RNA slot blots and Northern blots were used to characterize transcripts encoding ORFI. A transcript initiated 112 nt from the ilvGp2 promoter, but proceeding in the opposite direction, may encode the ORFI peptide. |
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| Keywords |
Amino Acid Sequence; Bacterial Proteins/genetics; Base Sequence; Blotting, Northern; Chromosome Mapping; Chromosomes, Bacterial; Codon; Escherichia coli/genetics; Escherichia coli Proteins; Isoleucine/biosynthesis; Molecular Sequence Data; Open Reading Frames; Restriction Mapping; Transcription, Genetic; Valine/biosynthesis |
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