title: Cloning and sequence analysis of Bacillus thuringiensis gene fragments isolated by restriction digest PCR.
abstract: OBJECTIVE
  To clone and analyze Bacillus thuringiensis gene fragments isolated by restriction digest PCR (RD-PCR).
  
  
  METHODS
  Specific primers were designed to amplify the genes of Bacillus thuringiensis israelensis (Bti), and the PCR products were classified and re-amplified by RD-PCR to obtain the fragments for subsequent purification and cloning into the pMD18-T vectors, followed by rapid identification. The recombinant plasmids were extracted from positive clones and the target gene fragments were sequenced.
  
  
  RESULTS
  Sequence analysis showed that all the fragments amplified were Bti genes.
  
  
  CONCLUSIONS
  RD-PCR is reliable in breaking down large gene fragments into confined and shorter gene fragments for preparing microarray probes.
authors.affiliation: Institute of Molecular Biochemistry, First Military Medical University, Guangzhou 510515, China. Wenli@fimmu.com
authors.forename: Xia
authors.initials: X
authors.lastname: Li
authors.forename: Wen-li
authors.initials: WL
authors.lastname: Ma
authors.forename: Yi
authors.initials: Y
authors.lastname: Pang
authors.forename: Bao
authors.initials: B
authors.lastname: Zhang
authors.forename: Li
authors.initials: L
authors.lastname: Jiang
authors.forename: Wen-Ling
authors.initials: WL
authors.lastname: Zheng
mesh_terms: D001413:Bacillus thuringiensis / Q000235:genetics*; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000032:analysis*; D005798:Genes, Bacterial; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA
chemical_list: D004269:DNA, Bacterial