A facile analytical method for the identification of protease gene profiles from Bacillus thuringiensis strains.
Five pairs of degenerate universal primers have been designed to identify the general protease gene profiles from some distinct Bacillus thuringiensis strains. Based on the PCR amplification patterns and DNA sequences of the cloned fragments, it was noted that the protease gene profiles of the three distinct strains of B. thuringiensis subsp. kurstaki HD73, tenebrionis and israelensis T14001 are varied. Seven protease genes, neutral protease B (nprB), intracellular serine protease A (ispA), extracellular serine protease (vpr), envelope-associated protease (prtH), neutral protease F (nprF), thermostable alkaline serine protease and alkaline serine protease (aprS), with known functions were identified from three distinct B. thuringiensis strains. In addition, five DNA sequences with unknown functions were also identified by this facile analytical method. However, based on the alignment of the derived protein sequences with the protein domain database, it suggested that at least one of these unknown genes, yunA, might be highly protease-related. Thus, the proposed PCR-mediated amplification design could be a facile method for identifying the protease gene profiles as well as for detecting novel protease genes of the B. thuringiensis strains.
Chen
Fu-Chu
FC
Institute of Biochemistry, National Yang Ming University, No. 155, Li-Nong St., Sec. 2, Peitou, Taipei 11221, Taiwan, ROC.
Shen
Li-Fen
LF
Chak
Kin-Fu
KF
D000595:Amino Acid Sequence; D001413:Bacillus thuringiensis / Q000201:enzymology* / Q000235:genetics*; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics*; D003001:Cloning, Molecular; D016384:Consensus Sequence; D017931:DNA Primers / Q000737:chemistry / Q000235:genetics; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D010450:Endopeptidases / Q000737:chemistry / Q000235:genetics*; D020869:Gene Expression Profiling; D016133:Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA
D001426:Bacterial Proteins; D017931:DNA Primers; D004269:DNA, Bacterial; D010450:Endopeptidases