Biochemical Investigation of Phenotypic Variance in Bdellovibrio bacteriovorus 109J at High Cell Density

Abstract

Preliminary data regarding the biochemical investigation of phenotypic variance in Bdellovibrio bacteriovorus 109J is presented. Host Independent B. bacteriovorus (HI-B) was successfully "flipped" from Host Dependent B. bacteriovorus (HD-B) by isolating HD-B prey lysate on a 0.45 ?m sterile polycarbonate filter suspended on a nutrient-rich LB agar plate. DNA from "Flipped" Host-Independent B. bacteriovorus 109J was isolated using a QIAgen DNeasy? Blood & Tissue Kit. Successful Polymerase Chain Reaction (PCR) was applied with primers designed with BLAST analysis, amplifying segments of the following genes in 109J: Bd2269 (putative serine protease), Bd0108 (hit locus), and Bd1626 (isopentenyl pyrophosphate isomerase). The genes of interest were visualized with Gel Electrophoresis, where E. coli prey presence did not affect the gene amplification of host-independent B. bacteriovorus . Real-time Polymerase Chain reaction will be applied in a future experiment to monitor relative gene expression of Bd1626 and other genes related to downstream isoprenoid synthesis in HD vs. HI- B. bacteriovorus to gain a greater molecular understanding of isoprenoid biosynthesis in B. bacteriovorus