Nucleotide sequence of the gene, protein purification and characterization of the pSC101-encoded tetracycline resistance-gene-repressor

Unger, B.; Becker, Jörn; Hillen, Wolfgang

doi:10.1016/0378-1119(84)90199-9

Cited by 26 publications

(21 citation statements)

References 25 publications

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“…Transcription of tet is initiated at nucleotide 45 (26); translation probably starts at position 86 and ends at 1273 (25). The tet gene comes from pSC101 (24), and its expression on that plasmid is induced by tetracycline (27). On pBR322, however, tet is expressed constitutively because the plasmid lacks the gene encoding the tet repressor (24,27).…”

Section: Resultsmentioning

confidence: 99%

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Topoisomerase I mutants: the gene on pBR322 that encodes resistance to tetracycline affects plasmid DNA supercoiling.

Pruss¹,

Drlica²

1986

Proc. Natl. Acad. Sci. U.S.A.

172

127

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Section: Resultsmentioning

confidence: 99%

“…The tet gene comes from pSC101 (24), and its expression on that plasmid is induced by tetracycline (27). On pBR322, however, tet is expressed constitutively because the plasmid lacks the gene encoding the tet repressor (24,27).…”

Section: Resultsmentioning

confidence: 99%

Topoisomerase I mutants: the gene on pBR322 that encodes resistance to tetracycline affects plasmid DNA supercoiling.

Pruss¹,

Drlica²

1986

Proc. Natl. Acad. Sci. U.S.A.

172

127

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“…Figure 4 shows an analysis of the cell associated proteins of E. coli WH1372 and E. coli WH602 at nine hours after induction. E. coli WH602 produces the same amount of a cytoplasmic protein, the pSC11 encoded Tet repressor, at the same time after induction from the same genetic background (16) Figure 4. The clear difference in the protein patterns of the cell fraction and the culture medium indicates, that disruption of cells cannot be the cause for the occurence of mutarotase in the culture medium.…”

Section: Analysis Of the Leader Peptidementioning

confidence: 99%

“…coli WH602 is E. coli K12 AHI t (15) transformed with pWH602 (16). pUR250 (17) was used to subclone a RsaI fragment to facilitate sequencing.…”

mentioning

confidence: 99%

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Acinetobacter calcoaceticusencoded mutarotase: nucleotide sequence analysis of the gene and characterization of its secretion inEscherichia coli

Gatz

Hillen

1986

Nucl Acids Res

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A threonine to alanine exchange at position 40 of Tet repressor alters the recognition of the sixth base pair of tet operator from GC to AT.

et al. 1988

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The tet operators of two naturally evolved tetracycline resistance determinants differ by a G.C to A.T transition at the sixth base pair. This mutation prevents heterologous recognition of these tet operators by their respective two Tet repressor proteins. The amino acid side chains responsible for this sequence‐specific distinction of operators were determined. For this purpose in vitro recombinants of the two tetR genes were constructed. Restriction sites were introduced by oligonucleotide‐directed mutagenesis in both genes followed by the exchange of different coding segments between them. The encoded chimeric Tet repressor proteins were expressed and their operator recognition specificity was scored in vivo. Exchanging gradually smaller coding segments led finally to a single amino acid exchange in both genes at position 40 of the primary structures. Each Tet repressor containing Thr at this position recognizes the G.C operator while those with Ala recognize the A.T operator regardless of the rest of the sequences. This result demonstrates clearly that the amino acid 40 of Tet repressor contacts and recognizes base pair 6 of tet operator. Sterical interference of the large Thr side chain with the methyl group of A.T and a possible involvement of the hydroxyl in hydrogen bonding to the operator are discussed as the molecular basis of this differentiation between A.T and G.C base pairs.

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Nucleotide sequence of the gene, protein purification and characterization of the pSC101-encoded tetracycline resistance-gene-repressor

Cited by 26 publications

References 25 publications

Topoisomerase I mutants: the gene on pBR322 that encodes resistance to tetracycline affects plasmid DNA supercoiling.

Topoisomerase I mutants: the gene on pBR322 that encodes resistance to tetracycline affects plasmid DNA supercoiling.

Acinetobacter calcoaceticusencoded mutarotase: nucleotide sequence analysis of the gene and characterization of its secretion inEscherichia coli

A threonine to alanine exchange at position 40 of Tet repressor alters the recognition of the sixth base pair of tet operator from GC to AT.

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