[Preparation of gene chip probes for Bacillus anthracis protective antigen].
OBJECTIVE
To study the method for rapid preparation of the gene chip probes for Bacillus anthracis protective antigen (pag).


METHODS
According to the sequences of pag published on PubMed, a pair of primers was designed to amplify the PA gene of Bacillus anthracis. Restriction enzyme digestion and AT clone were employed to rapidly screen out the recons of PA segments for the preparation of the gene chip probes. The DNA sequences of these segments were determined with 377 DNA Sequencer, and the resultant sequences analyzed with Bioinformatic software.


RESULTS
With the designed primers, the pag 2,205 bp in length was amplified. After enzyme digestion and AT clone, 7 fragments of varied lengths were screened, which underwent analysis with the basic local alignment sequence tool (BLAST) and 377 DNA Sequencer. BLAST analysis showed that all the fragments cloned belonged to Bacillus anthracis.


CONCLUSIONS
PCR in conjunction with restriction enzyme digestion and AT clone is rapid and simple to prepare the desired gene chip probes.
Ma
Xiao-dong
XD
Institute of Molecular Biology, First Military Medical University, Guangzhou 510515, China.
Ma
Wen-li
WL
Lü
Liang
L
Xiao
Wei-wei
WW
Sun
Zhao-hui
ZH
Zheng
Wen-ling
WL
D000942:Antigens, Bacterial; D001408:Bacillus anthracis / Q000276:immunology*; D001427:Bacterial Toxins / Q000235:genetics*; D001483:Base Sequence; D008969:Molecular Sequence Data; D020411:Oligonucleotide Array Sequence Analysis; D016133:Polymerase Chain Reaction
D000942:Antigens, Bacterial; D001427:Bacterial Toxins; C030325:anthrax toxin