问号钩端螺旋体基因组DNA芯片的研制

Construction and validation of Leptospira interrogans genome DNA microarray

目的研制并初步评估问号钩端螺旋体(简称钩体)赖型赖株的基因组DNA芯片。方法利用Primegens引物设计软件筛选出问号钩体赖型赖株全基因组中的特异性基因进行引物设计。对成功设计出相应引物的3 290个基因用聚合酶链反应方法进行扩增,以纯化后的产物点样制备芯片。并用双色荧光杂交策略对芯片质量进行了初步平估。结果共获得3 290个基因产物用于点样。参考株自身杂交实验结果表明:该芯片有较高的点一致性、信噪比和较低的假阳性率。结论成功制备了包含问号钩体赖型赖株3 290个目的基因的基因组DNA芯片,并可用于基于该芯片的问号钩体比较基因组学的研究。

Objective To develop and validate a genomie DNA microarray of L. irtterrogans serovar Lai str. 56601.Methods Primers were designed based on specific coding gene sequences of L. interrogans serovar Lai str. 56601 using a computer seftware Primegens. PCR products from these genes ,sere purified and printed onto the glass sides in triplicate. The qualit＇y of this miemarray is evaluated bv the method of two-fluoreseenee comparative hybridization. Results A total of 3 290 genes were successfully anlplified by PCR. Genomie DNAs from an autologous reference strain were labeled with Cy3 or Cy5 fluor and the labeled probes were hybridized against this new microarray. Results showed high consistency of spots, high signal/noise ratio and low false positive rate. Conclusion A mieroarray was developed contailfing 3 290 ulfique gene sequences from L. interrogans serovar Lai str. 56601. A reserch ptatform bassed on this micoarray can be further established to conduct comparative genomic analysis for L. interrogans.