Centro de Estudios de Biodiversidad y Biotecnología, Centro de Investigaciones Biológicas, Fundación para Investigaciones Biológicas Aplicadas (CEBB-CIB FIBA), C.C. 1348, 7600, Mar del Plata, Argentina.
Curr Microbiol. 2011 Mar;62(3):866-70. doi: 10.1007/s00284-010-9776-1. Epub 2010 Nov 4.
PCR has been widely used to identify cry-type genes, to determine their distribution, to detect new such genes and to predict insecticidal activities. We describe here a molecular approach to analyze the genetic diversity of B. thuringiensis cry-like genes based on denaturing gradient gel electrophoresis (DGGE). This analysis demonstrated that different B. thuringiensis isolates can be distinguished according to its PCR-DGGE profile of cry-like genes. Identification of the resolvable DNA fragments was easy to accomplish by DNA sequencing, which was confirmed in this work. Importantly, the strategy allowed the identification of unknown B. thuringiensis cry-like sequences present in a single strain that remained cryptic after PCR analysis using degenerate primers. The method developed in this work contributes to the availability of molecular techniques for both B. thuringiensis strains and cry-like genes identification and discovery.
PCR 已被广泛用于鉴定 cry 型基因，以确定其分布，检测新的此类基因，并预测杀虫活性。我们在这里描述了一种基于变性梯度凝胶电泳（DGGE）分析苏云金芽孢杆菌 cry 样基因遗传多样性的分子方法。该分析表明，根据 cry 样基因的 PCR-DGGE 图谱，可以区分不同的苏云金芽孢杆菌分离株。通过 DNA 测序很容易鉴定可分辨的 DNA 片段，这在本工作中得到了证实。重要的是，该策略允许鉴定在使用简并引物进行 PCR 分析后仍保持隐匿状态的单个菌株中存在的未知苏云金芽孢杆菌 cry 样序列。本工作中开发的方法有助于获得分子技术，用于鉴定和发现苏云金芽孢杆菌菌株和 cry 样基因。
