CLONING AND SEQUENCING OF A HEMAGGLUTININ COMPONENT OF THE BOTULINUM NEUROTOXIN COMPLEX ENCODED BY CLOSTRIDIUM-BOTULINUM TYPE-A AND TYPE-B

EAST AK STACEY JM COLLINS MD

Ak. East et al., CLONING AND SEQUENCING OF A HEMAGGLUTININ COMPONENT OF THE BOTULINUM NEUROTOXIN COMPLEX ENCODED BY CLOSTRIDIUM-BOTULINUM TYPE-A AND TYPE-B, Systematic and applied microbiology, 17(3), 1994, pp. 306-312

15

INGLESE

Article

Microbiology,"Biothechnology & Applied Migrobiology

Systematic and applied microbiology → ACNP

0723-2020

17

3

1994

306 - 312

ISI

0723-2020(1994)17:3<306:CASOAH>2.0.ZU;2-6

Degenerate oligonucleotide primers designed to the N-terminal amino ac id sequence of proteins purified from the botulinum neurotoxin complex were used to amplify DNA fragments using PCR from genomic DNA of Clos tridium botulinum types A: NCTC 7272, and B: NCTC 7273 (proteolytic st rain) and Eklund 17B (non-proteolytic strain). A fragment of similar t o 1.9 kb was amplified using template DNA from each strain, which was subsequently cloned in E. coli and the sequences determined. Two open reading frames (orfs) were discovered: one corresponding to a protein similar to 33.7 kD which shows between 36 and 41% identity with the ma jor hemagglutinin component (HA-33) of the botulinum progenitor comple x encoded by C. botulinum type C. The second orf, encoding a protein o f similar to 21.7 kD, P-21, of 178 amino acids (179 in strain NCTC 727 3) has similar to 93% identity between the type A and two type B strai ns, but is absent from type C.