Restriction enzyme analysis and DNA sequencing comparison for alpha-toxin gene among different types of Clostridium perfringens.
Alpha-toxin is produced by all types of Clostridium perfringens. The genes encoding alpha-toxin from the available five types of Clostridium perfringens [A (chicken strain), A (rabbit strain), B, C and D] were PCR amplified using specific primers and the PCR products were examined on 1.5% (w/v) agarose gel and demonstrated the same bands comparable to the published alpha-toxin gene. Restriction enzyme analysis using two sets of enzymes (one set known to have recognition sites; Hinf1, EcoRV and Mse1 and the other set known to lack recognition sites Hind III, Pst1 and BamH1) were carried out. The first set of enzymes revealed the same cut specific for alpha-toxin gene. However, the second set of enzymes revealed no cut which is consistent to the published data. The PCR products of alpha-toxin gene from the five types were separately sequenced and aligned with all published alpha-toxin genes of Clostridium perfringens. Identities among all studied alpha-toxin gene sequences and with the published ones were nearly 96-98%. There are no any significant differences among these nucleotide sequences. It is concluded that alpha-toxin gene sequences among different types of Clostridium perfringens are similar and highly conserved.
Effat
M M
MM
Department of Microbiology and Immunology, National Research Centre, Giza, Egypt.
D001427:Bacterial Toxins / Q000235:genetics*; D001483:Base Sequence; D002135:Calcium-Binding Proteins / Q000235:genetics*; D003016:Clostridium perfringens / Q000235:genetics*; D004269:DNA, Bacterial / Q000235:genetics*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012689:Sequence Homology, Nucleic Acid; D010738:Type C Phospholipases / Q000235:genetics*
D001427:Bacterial Toxins; D002135:Calcium-Binding Proteins; D004269:DNA, Bacterial; D010738:Type C Phospholipases; C040649:alpha toxin, Clostridium perfringens