炭疽杆菌保护性抗原基因芯片探针的制备

目的快速制备炭疽杆菌保护性抗原基因芯片探针。方法根据PubMed上公布的炭疽杆菌保护性抗原的DNA序列设计引物,扩增保护性抗原基因。利用酶切、AT克隆方法快速分析筛选出保护性抗原基因片段的重组子,从而制备成芯片探针。DNA自动分析仪对克隆片段进行序列测定,生物信息学软件对其基因序列进行分析。结果根据炭疽杆菌保护性抗原设计的引物,可以扩增出长2 205 bp的保护性抗原基因,经酶切、AT克隆方法筛选出约7个长度不一的片段,对其片段进行序列测定并进行Blast序列比对得知这些片段均属于炭疽杆菌保护性抗原基因。结论利用PCR扩增产物并结合酶切、AT克隆方法可以快速、简便地制备基因芯片探针。

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. Conclusion PCR in conjunction with restriction enzyme digestion and AT clone is rapid and simple to prepare the desired gene chip probes.