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Publisher Summary The chapter describes Borrelia burgdorferi membrane arrays contain PCR-amplified open reading frames (ORFs) from Borrelia burgdorferi strain B31 MI, and illustrates the strain whose genome sequence. The B. burgdorferi B31 genome is unique among fully sequenced bacterial genomes in that it consists of a linear chromosome 910,725 bp in length and a collection of 9 circular and 12 linear plasmids. Among the 1689 putative open reading frames (ORFs), 855 are chromosome encoded and 834 are plasmid encoded. More than 90% of the Borrelia burgdorferi plasmid ORFs are unrelated to any known bacterial sequences. The novel genes found on the Borrelia burgdorferi plasmids may, therefore, contribute to the ability of this pathogen to survive and maintain its complex life cycle. Since the identities of all the spots (ORFs) on the array are known, the complete gene expression profile of an organism under a given set of conditions may be analyzed with a single array.

Publisher Summary Spirochaetes are a phylogenetically ancient bacterial group and one of the few for which classification by morphology is consistent with molecular typing based upon rRNA sequence comparisons. Although a unique spiral shape and periplasmic flagella for motility are universal among the spirochaetes, they are a diverse group of bacteria with regard to many other characteristics. Several spirochaetes represent important human and animal pathogens, with histories of disease that extend for centuries. None of the spirochaetes has been the subject of extensive genetic analysis and there is a corresponding paucity in current genetic tools and methods with which to manipulate them. All other buffers used for analytical purposes such as gel electrophoresis are prepared with deionized water. Instead of the water drawn from a Millipore system, double-distilled water can be used as well. Media for cultivating and storing bacteria generally include liquid media, solid media, storage media, and supplements. Solid media are mostly liquid media solidified by the addition of agar.

The focus of research in our laboratory over the few years has been the regulation of synthesis, processing, and release of the ACTH/LPH family of peptides. These peptides are derived from a common precursor protein that is found in both the anterior and intermediate lobes of the pituitary (Roberts et al., 1978) and in the hypothalamus (Liotta et al., 1979). In the anterior lobe this protein gives rise to α (1–39) ACTH, β-lipotropin (β-LPH) and an N-terminal fragment of undefined function. In addition, a variety of intermediate lobe pituitary peptides can be derived from the precursor by further processing of ACTH and β-LPH. In this paper we compare the structure of the precursor in the anterior and intermediate lobes of mouse and rat pituitary. Processing of the precursor to its constituent hormones is then contrasted in primary cultures of anterior and intermediate lobe cells using pulse label and pulse chase techniques with radioactive amino acids and sugars.