Identification and characterization of a polysaccharide deacetylase gene from Bacillus thuringiensis.
One polysaccharide deacetylase gene was cloned from Bacillus thuringiensis and designated pdaA. Disruption of pdaA did not affect vegetative growth and sporulation but obviously affected spore germination. When L-alanine was added into the spore suspension, the spores of the pdaA disruption mutant showed a slow and partial reduction in absorbance at OD600 and became phase pale gray compared with phase dark of the wild-type strain. In contrast with the outgrowing of wild-type spores after germination, the pdaA mutant spores were blocked at the stage of spore germination. Transmission electron micrographs revealed a significant difference between the pdaA mutant and the wild-type strain in the spore cortex. Introduction of the pdaA gene into the pdaA disruption mutant complemented the germination-negative phenotype. Reverse transcription--polymerase chain reaction showed that pdaA was transcribed after incubation for 10 h in CCY medium.
Hu
Kun
K
Institute of Microbiology, Chinese Academy of Science, Beijing, PR China.
Yang
Haihua
H
Liu
Gang
G
Tan
Huarong
H
D000581:Amidohydrolases / Q000235:genetics*; D001413:Bacillus thuringiensis / Q000201:enzymology* / Q000235:genetics* / Q000502:physiology; D003001:Cloning, Molecular; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D046529:Microscopy, Electron, Transmission; D009154:Mutation; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013171:Spores, Bacterial / Q000235:genetics / Q000502:physiology / Q000648:ultrastructure; D014158:Transcription, Genetic
D000581:Amidohydrolases; C022427:polysaccharide deacetylase