title: [Establishment of the genotype-specific targets of botulinum neurotoxins types B and E based on reverse genetics].
abstract: OBJECTIVE
  To establish the genotype-specific targets plasmids and engineered E.coli strains of botulinum neurotoxins (BoNT) types B and E based on reverse genetics.
  
  
  METHODS
  The gene sequences of BoNT were obtained from GenBank and analyzed using DNAMAN, Lasergene, Vector NTI and BLAST. Two target fragments of BoNT/B and BoNT/E were anchored and then synthesized as 5 and 10 short DNA single strands, respectively. The full-length target sequences were amplified by overlapping PCR and subcloned into pMD 18-T vector, and the recombinant plasmids were identified by restriction enzyme digestion and sequencing.
  
  
  RESULTS
  Sixty full-length sequences of 4 types of BoNT, namely types A, B, E, and F, were available in GenBank. Two target fragments, BoNT/B of 215 bp and BoNT/E of 360 bp, and their specific primer pairs were anchored after sequence analysis. pMD 18-T-BoNT/B and pMD 18-T-BoNT/E containing these two target sequences were confirmed.
  
  
  CONCLUSIONS
  The engineered plasmids and E.coli stains containing the genotype-specific target fragments of BoNT/B and BoNT/E have been constructed successfully.
authors.affiliation: Department of Parasitology, School of Public Health and Tropical Medicine, NanFang Hospital Southern Medical University, Guangzhou, China. liying20920@hotmail.com
authors.forename: Ying
authors.initials: Y
authors.lastname: Li
authors.forename: Chun-li
authors.initials: CL
authors.lastname: Yang
authors.forename: Ying
authors.initials: Y
authors.lastname: Zhao
authors.forename: Yu-qi
authors.initials: YQ
authors.lastname: Guo
authors.forename: Chang-zheng
authors.initials: CZ
authors.lastname: Liu
authors.forename: Xiao-hong
authors.initials: XH
authors.lastname: Zhou