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Journal of Applied Sciences
Year: 2006  |  Volume: 6  |  Issue: 7  |  Page No.: 1540 - 1546

Differentiation of Bacillus thuringiensis and Escherichia coli by the Randomly Amplified Polymorphic DNA Analysis

Halima Hassan Salem, Tian-Hua Huang, Bahy Ahmed Ali and Qing-Dong Xie    

Abstract: Random Amplified Polymorphic DNA (RAPD) is a method of producing a biochemical fingerprint of a particular species. This study involved the isolation of DNA from three Bacillus thuringiensis (B. thuringiensis) [B. thuringiensis tenebrionis (tt4) and B. thuringiensis israelensis (i977)] as reference strains and new isolate B. thuringiensis strain (66) using the DNeasy Mini Kit (Qiagen, Inc. protocol). Two Escherichia coli (E. coli BL21 and E. coli C1a) DNAs are provided as components of RAPD Analysis Kit. RAPD analysis was performed using Ready-To-GoTM Analysis Beads and nine primers. Bands were then analyzed using a binary system and a phylogenetic tree created. The five strains segregated into two major clusters. The maximum linkage distance between the two major clusters was 103 units. First cluster consisted of two E. coli strains and the second cluster consisted of the three B. thuringiensis strains based on genetic similarity in their RAPD profiles. The B. thuringiensis reference strains were located in subcluster showing high similarity. Genetic similarities estimated as band sharing were calculated and all comparisons among bacterial strains have been done. The results showed that RAPD-PCR is powerful method for differentiation between B. thuringiensis and E. coli.

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