Synthesis and Cloning of a Small Bacillus Pheromone Gene (ComXRO-B-2) by Primer-Dimer Formation with PCR

Authors : Devrim Demir Dora, Tanıl Kocagöz, Filiz Öner

Pages : 765-771

View : 14 | Download : 7

Publication Date : 9999-12-31

Article Type : Makaleler

Abstract :ComX pheromone is the major extracellular signaling peptide stimulating transformation in response to high cell density in Bacillus species. In this study, Bacillus mojavensis ComX(RO-B-2) pheromone gene was cloned to pGEMT-Easy vector based on TA cloning of PCR products. The gene encoding 11 amino acid peptides of Bacillus mojavensis RO-B-2 strain ComX pheromone (GLQIYTNWVPS) was obtained by PCR amplification of 2 primers complementary to each other at their 3' end. This eliminated the need for the original bacterial gene as DNA template for PCR. The amplified PCR products were ligated directly without any modification by T4 DNA ligase into pGEMT-Easy vector, which has a single overhanging T residue at the 3' ends of the cloning site.
Keywords : Bacillus mojavensis RO-B-2, ComX pheromone gene, TA cloning, primer-dimer, PCR.