Streptomyces aureofaciens B96 produces several intra- and extracellular enzymes with deoxyribonuclease activity. According to the sequence of the previously published gene exoSc from S. coelicolor A3(2), the DNA sequence from S. aureofaciens B96 was amplified, cloned and expressed in E. coli. The protein product of exoSa gene, recExoSa, was also an exonuclease with DNAase and 5'-phosphomonoesterase activities at optimum temperature 37 degrees C and pH 8.0. It degraded only linear DNA (chromosomal, double-stranded and single-stranded) and linear plasmid DNA from both ends, with a preference to blunt ends in comparison with overhang ends. The purified enzyme exhibited no RNAase activity. Both exoSc and exoSa genes were interrupted by the apramycin resistance gene; constructed fragments were transformed into particular streptomyces protoplasts. Mutation caused by exoSa disruption in S. aureofaciens chromosome and mutation by interrupted exoSc in S. coelicolor were lethal.
- MeSH
- bakteriální proteiny MeSH
- deoxyribonukleasy MeSH
- exonukleasy MeSH
- financování organizované MeSH
- fosfatasy MeSH
- klonování DNA MeSH
- molekulární sekvence - údaje MeSH
- sekvence aminokyselin MeSH
- sekvenční seřazení MeSH
- stabilita enzymů MeSH
- Streptomyces aureofaciens enzymologie genetika chemie MeSH
- Streptomyces coelicolor enzymologie genetika chemie MeSH
- substrátová specifita MeSH
- umlčování genů MeSH
A multiplex PCR method was developed to characterize the potential variability within the sip gene from bovine isolates of Streptococcus agalactiae. An incomplete sip gene (ncosip) was found in four isolates of S. agalactiae. According to the in silico analysis of the missing region at amino acid level, the N-terminal of surface immunogenic protein (Sip) was found to contain a LysM domain motif; whilst the incomplete Sip (NcoSip) lacked a part of this motif. Immunity elicitation against Sip was confirmed by immunization of mice. This response was partially observed also with NcoSip.
- MeSH
- aminokyselinové motivy MeSH
- antigeny bakteriální genetika chemie imunologie MeSH
- financování organizované MeSH
- lidé MeSH
- molekulární sekvence - údaje MeSH
- myši inbrední BALB C MeSH
- myši MeSH
- protilátky bakteriální krev MeSH
- sekvence aminokyselin MeSH
- Streptococcus agalactiae genetika chemie imunologie MeSH
- streptokokové infekce imunologie mikrobiologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- myši MeSH
- zvířata MeSH
- MeSH
- DNA restrikčně-modifikační enzymy izolace a purifikace klasifikace MeSH
- finanční podpora výzkumu jako téma MeSH
- hospodářská zvířata mikrobiologie MeSH
- plazmidy diagnostické užití izolace a purifikace MeSH
- restrikční enzymy izolace a purifikace klasifikace MeSH
- Staphylococcus aureus enzymologie MeSH
- Streptococcus agalactiae enzymologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
