BOX-PCR技术在微生物多样性研究中的应用

Applications of BOX-PCR Technique in Diversity Study of Microorganisms

摘要:

近年来在微生物多样性研究中, 利用微生物基因组中广泛分布的短重复序列设计引物, 选择性地扩增重复序列之间的不同基因区域, 以得到大小不等的DNA扩增片段的方法日渐增多。以BOX插入因子(细菌基因组重复序列)为基础的PCR技术, 具有操作简单快捷, 可重复性强, 容易获得较为丰富的扩增条带等特点, 最初主要应用于细菌的多样性研究。目前研究发现用BoxA1R引物对微生物中的真菌、放线菌进行选择性的扩增, 也能够达到很好的遗传及多样性分析的目的。本文综述了BOX-PCR指纹图谱分析技术的特点和一般步骤; 结合作者对植物内生细菌的BOX-PCR指纹图谱分析体系的优化, 对BOX-PCR技术的改进进行了总结; 并对该技术在微生物菌株多样性研究领域的应用现状和前景进行了阐述。

Abstract:

In recent years, the molecular methods using short tandem repeats sequences in microbial genome to design primers and amplify the sequences have been continuously reported. The PCR based on the BOX elements is a simple, fast, highly reproducible method for comparing patterns of the amplified bands, and it was applied in studies on the classification of bacteria originally. Researchers found that it could be used as well for analysis of the genetic diversities of fungi and streptomyces using BOXA1R primer to amplify the BOX elements. This paper reviewed the characteristics and the general procedure of BOX-PCR fingerprinting. The system optimization of BOX-PCR fingerprinting for endophytic bacteria and BOX-PCR technology improvements are summarized. Its applications and prospect on diversity research of different microorganisms are discussed as well.