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Nucleotide sequence and expression of the Pseudomonas aeruginosa algF gene controlling acetylation of alginate
Author(s) -
Shinabarger Dean,
May Thomas B.,
Boyd Aoife,
Ghosh Mita,
Chakrabarty A. M.
Publication year - 1993
Publication title -
molecular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.857
H-Index - 247
eISSN - 1365-2958
pISSN - 0950-382X
DOI - 10.1111/j.1365-2958.1993.tb01232.x
Subject(s) - biology , acetylation , mutant , gene , nucleic acid sequence , pseudomonas aeruginosa , plasmid , microbiology and biotechnology , gene expression , peptide sequence , biochemistry , pseudomonas , nucleotide , pseudomonadaceae , bacteria , genetics
Summary Colonization of the cystic fibrosis lung by Pseudomonas aeruginosa is greatly facilitated by the production of an exopolysaccharide called alginate. In this study we determined the nucleotide sequence of an alginate modification gene, algF , which controls the addition of acetyl groups to alginate. Expression of algF using a T7 promoter‐expression system showed that algF codes for a 24.5 kDa polypeptide (predicted size 22 832 Da) that is processed to 19.5 kDa. The N ‐terminus of the processed polypeptide matched the predicted amino acid sequence of AlgF starting at Asp‐29. An algF mutant failed to produce alginate owing to a polar effect on the downstream algA gene. Although the algA gene, provided in trans , restored synthesis of alginate, the alginate was non‐acetylated. We show that a plasmid containing both the algF and algA gene complements the alginate acetylation defect of the algF mutant strain.