Molecular cloning of the gene encoding the mosaic neurotoxin, composed of parts of botulinum neurotoxin types C1 and D, and PCR detection of this gene from Clostridium botulinum type C organisms.

Applied and Environmental Microbiology
K MoriishiK Oguma

Abstract

The DNA fragment common to the genes encoding botulinum neurotoxin types C1 (BN/C1) and D (BN/D) was amplified by PCR from the culture supernatant of Clostridium botulinum type C strain 6813 (C6813) that was treated with either DNase I or proteinase K but not from the supernatant that was treated with both DNase I and proteinase K, suggesting the neurotoxin gene is located on a certain bacteriophage DNA. Thus, to isolate the neurotoxin gene, we performed PCR with the culture supernatant of C6813 and seven primer pairs designed from the genes encoding BN/C1 and BN/D. The coding region in the connected sequence encodes a neurotoxin composed of 1,280 amino acids with a molecular weight of 147,817. The neurotoxin from C6813 has 95% amino acid identity to BN/C1, except for its C-terminal one-third, which is quite similar to the C-terminal one-third of BN/D (95% identity). When we performed PCRs with four primer pairs designed from the 5'-terminal two-thirds of the BN/C1 gene and two primers from the 3'-terminal one-third of the BN/D gene, DNA fragments of the expected sizes (0.5 to 1.3 kbp) could be amplified from C. botulinum type C strains 6812 and 6814. These results suggest that some strains of C. botulinum type C contain the ge...Continue Reading

References

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Citations

Jan 8, 2020·Toxins·Christine Rasetti-EscargueilMichel R Popoff
Jun 15, 2007·Critical Reviews in Microbiology·Shuowei CaiShashi Sharma
Apr 4, 2000·Physiological Reviews·G SchiavoC Montecucco
May 12, 2018·Toxins·Domenico Azarnia Tehran, Marco Pirazzini
Oct 9, 2019·Antimicrobial Agents and Chemotherapy·Doris M SnowMilan T Tomic
Oct 12, 2001·Toxicon : Official Journal of the International Society on Toxinology·E A Johnson, M Bradshaw
Nov 23, 2006·Journal of Bacteriology·K K HillJ D Marks

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