We have analysed the genes borne on a 6·0 kb Hin dIII fragment cloned from the chromosome of Clostridium botulinum type E strain Mashike. This fragment, cloned within plasmid pU9EMH, contains part of the structural gene for botulinum toxin type E neurotoxin as well as the entire structural gene for a nontoxic component of botulinum type E progenitor neurotoxin gene, ent-120. ent-120 is transcribed in the same direction as the neurotoxin gene and consists of one open reading frame encoding 1162 amino acid residues. Western blotting with anti-nontoxic component sera demonstrates that ent-120 encodes a protein of 120 kDa which forms part of the nontoxic component. ent-120 is homologous to an analogous gene found in botulinum type C strains (69.3% identity at the nucleotide level and 56.1% at the amino acid level). Two stretches of amino acids at the N-terminus of the ent-120 protein are highly homologous to amino acid sequences within the type E neurotoxin. The stop codon of the ent-120 gene is situated 27 nucleotides upstream from the start codon of the neurotoxin gene.
BinzT., KurazonoH., WilleM., FrevertJ., WernarsK., NiemannH.1990; The complete sequence of botulinum neurotoxin A and comparison with other clostridial neurotoxins. Journal of Biological Chemistry 265:9153–9158
FujiiN., KimuraK., MurakamiT., IndohT., YashikiT., TsuzukiK., YokosawaN., OgumaK.1990; The nucleotide and deduced amino acid sequences of EcoRl fragment containing the 5’-terminal region of Clostridium botulinum type E toxin gene cloned from Mashike, Iwanai and Otaru strains. Microbiology and Immunology 34:1041–1047
FujiiN., KimuraK., YashikiT., IndohT., MurakamiT., TsuzukiK., YokosawaN., OgumaK.1991; Cloning of a DNA fragment encoding the 5’-terminus of the botulinum type E toxin gene from Clostridium butyricum strain BL6340. Journal of General Microbiology 136:519–525
FujiiN., KimuraK., YashikiT., TsuzukiK., MoriishiK., YokosawaN., SyutoB., OgumaK.1992; Cloning and whole nucleotide sequence of the gene for the light chain component of botulinum type E toxin for Clostridium butyricum strain BL6340 and Clostridium botulinum type E strain Mashike. Microbiology and Immunology 36:213–220
KitamuraM., SakaguchiS., SakaguchiG.1968; Purification and some properties of Clostridium botulinum type E toxin. Biochimica et Biophysica Acta 168:207–217
KimuraK., FujiiN., TsuzukiK., MurakamiT., IndohT., YokosawaN., TakeshiK., SyutoB., OgumaK.1990; The complete nucleotide sequence of the gene coding for botulinum C1 toxin in the c-st phage genome. Biochemical and Biophysical Research Communications 171:1304–1311
KimuraK., FujiiN., TsuzukiK., YokosawaN., OgumaK.1992; The functional domains of Clostridium botulinum type C neurotoxin. In Recent Advances in Toxinology Research, pp. 375–385, Edited by. P. Gopalakrishnakone; Singapore: Venom and Toxin Research Group.:
PouletS., HauserD., QuanzM., NiemannH., PopoffM.R.1992; Sequences of the botulinal neurotoxin E derived from Clostridium botulinumtype E (strain Beluga) and Clostridium butyricum (strains ATCC 43181 and ATCC 43755). Biochemical and Biophysical Research Communications 183:107–113
SomersE., DasguptaB.R.1991; Clostridium botulinum types A, B, C, and E produce proteins with or without hemagglutinating activity: do they share common amino acid sequences and genes?. Journal of Protein Chemistry 10:415–425
ThompsonD.E., BrehmJ.K., OultramJ.D., SwinfieldT.J., ShoneC.C., AtkinsonT., MellingJ., MintonN.P.1990; The complete amino acid sequence of the Clostridium botulinum type A neurotoxin, deduced by nucleotide sequence analysis of the encoding gene. European Journal of Biochemistry 189:73–81
TsuzukiK., KimuraK., FujiiN., YokosawaN., IndohT., MurakamiT., OgumaK.1990; Cloning and complete nucleotide sequence of the gene for the main component of hemagglutinin produced by Clostridium botulinum type C. Infection and Immunity 58:3173–3177
TsuzukiK., KimuraK., FujiiN., YokosawaN., OgumaK.1992; The complete nucleotide sequence of the gene coding for the nontoxic-nonhemagglutinin component of Clostridium botulinum type C progenitor toxin. Biochemical and Biophysical Research Communications 183:1273–1279
WhelanS.M., ElmoreM.J., BodsworthN., BrehmJ.K., AtkinsonT., MintonN.P.1992; Molecular cloning of the Clostridium botulinum structural gene encoding the type B neurotoxin and determination of its entire nucleotide sequence. Applied and Environmental Microbiology 58:2345–2354
WhelanS.M., ElmoreM.J., BodsworthN.J., AtkinsonT., MintonN.P.1992b; The complete amino acid sequence of the Clostridium botulinum type E neurotoxin, derived by nucleotide- sequence analysis of the encoding gene. European Journal of Biochemistry 204:657–667
YokosawaN., TsuzukiK., SyutoB., OgumaK.1986; Activation of Clostridium botulinum type E toxin purified by two different procedures. Journal of General Microbiology 132:1981–1988