1992
DOI: 10.1128/jb.174.7.2208-2214.1992
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Staphylococcal phosphoenolpyruvate-dependent phosphotransferase system: molecular cloning and nucleotide sequence of the Staphylococcus carnosus ptsI gene and expression and complementation studies of the gene product

Abstract: A digoxigenin-labeled DNA probe that was complementary to the gene ptsH and the beginning of the gene ptsI was used to clone a 3.2-kb HincII-BamHI restriction fragment containing the complete ptsI gene of Staphylococcus carnosus. The restriction fragment was cloned in the antisense orientation to the lac promoter in the low-copy-number vector pSU18. The nucleotide sequences of the ptsI gene, which encodes enzyme I (EC 2.7.3.9), and the corresponding flanking regions were determined. The primary translation pro… Show more

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Cited by 29 publications
(23 citation statements)
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References 29 publications
(19 reference statements)
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“…The erythromycin (Em) resistance gene ermB was isolated from plasmid pI258 [14] and subsequently introduced in the plasmids pUC18, pUC19 or pBluescript KS + (Stratagene). Plasmid pUC19‐ ptsI served as the source of the phosphotransferase gene ptsI from S. carnosus [15]. The sucrose utilization plasmid pRA7 [16] was used to inactivate the scrB gene of S. xylosus .…”
Section: Methodsmentioning
confidence: 99%
“…The erythromycin (Em) resistance gene ermB was isolated from plasmid pI258 [14] and subsequently introduced in the plasmids pUC18, pUC19 or pBluescript KS + (Stratagene). Plasmid pUC19‐ ptsI served as the source of the phosphotransferase gene ptsI from S. carnosus [15]. The sucrose utilization plasmid pRA7 [16] was used to inactivate the scrB gene of S. xylosus .…”
Section: Methodsmentioning
confidence: 99%
“…Protein Expression and Purification-EI from S. carnosus was overexpressed in Escherichia coli (strain DH5␣) harboring the plasmid pUC-ptsO2.6X (23). Cells were grown in the presence of isopropyl-␤-D-thiogalactoside in TBY broth (10 g of Tryptone, 5 g of NaCl, 5 g of yeast extract/liter).…”
Section: Methodsmentioning
confidence: 99%
“…6 The atomic coordinates have been deposited with the Protein Data Bank (www.rcsb.org), under accession code 2HRO. (23) was overexpressed and purified as described under "Experimental Procedures." Initial hexagonal crystals appearing in Hampton Standard screens diffracted poorly, whereas monoclinic crystals, appearing 3-5 days later, diffracted up to 2.5 Å.…”
Section: Methodsmentioning
confidence: 99%
“…EI of the PTS was purified from Staphylococcus carnosus and HPr was purified from B. subtilis as described before (16,24). For the purification of LevR, the levR gene was cloned under the control of the strong degQ36 promoter (39).…”
Section: Methodsmentioning
confidence: 99%