Cloning and sequence analysis of Bacillus thuringiensis gene fragments isolated by restriction digest PCR.
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.
Li
Xia
X
Institute of Molecular Biochemistry, First Military Medical University, Guangzhou 510515, China. Wenli@fimmu.com
Ma
Wen-li
WL
Pang
Yi
Y
Zhang
Bao
B
Jiang
Li
L
Zheng
Wen-Ling
WL
D001413:Bacillus thuringiensis / Q000235:genetics*; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000032:analysis*; D005798:Genes, Bacterial; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA
D004269:DNA, Bacterial