Infect Immun 2001,69(7):4691–4694.CrossRefPubMed 44. Baron GS, Nano FE: MglA and MglB are required for the intramacrophage growth of Francisella novicida. Mol Microbiol 1998,29(1):247–259.CrossRefPubMed 45. Rueger B, Thalhammer J, Obermaier I, Gruenewald-Janho S: Experimental procedure for the detection of a rare human mRNA with the DIG System. Front selleckchem Biosci 1997, 2:c1–5.PubMed 46. Honeyman AL, Cote CK, Curtiss R 3rd: Construction of transcriptional and translational lacZ gene reporter plasmids for use in Streptococcus mutans. J Microbiol Methods 2002,49(2):163–171.CrossRefPubMed
47. LoVullo ED, Sherrill LA, Perez LL, Pavelka MS Jr: Genetic tools for highly Selleckchem Z-DEVD-FMK pathogenic Francisella tularensis subsp. tularensis. Microbiology 2006,152(Pt 11):3425–3435.CrossRefPubMed 48. Horton RM, Hunt HD, Ho SN, Pullen JK, Pease LR: Engineering
hybrid genes without the use of restriction enzymes: gene splicing by overlap extension. Gene 1989,77(1):61–68.CrossRefPubMed 49. Miller JH: Experiments in molecular genetics. Cold Spring Harbor, N.Y.: Cold Spring Harbor Laboratory 1972. Authors’ contributions JF carried out all experiments with the participation of TMK and SB in the extracellular galactosidase assays. TMK and SB helped draft the manuscript and provided intellectual input to data analysis. THK and JF designed and coordinated experiment, analyzed data, and drafted the manuscript. All authors read and approved the final
manuscript.”
“Background Contagious bovine pleuropneumonia (CBPP), a pulmonary disease caused by Temsirolimus Mycoplasma mycoides subsp. mycoides SC (MmmSC) is a major constraint to cattle production P-type ATPase in Africa [1]. The current vaccines are not always fully effective [2] and there remains an urgent need to control or even eradicate the disease. Although the nucleotide sequence of the MmmSC type strain PG1 genome is available, the proteins responsible for protection have not been identified. Accordingly, an important step towards a subunit vaccine would be to identify which of the potentially large number of antigens encoded in its genome [3–5] actually trigger immune responses during infection. Serum antibodies are likely to be involved in immunity since passive transfer of sera from recovered cattle can protect recipient calves [6, 7], but Th1 memory lymphocytes and γδ T-cells are also active [8–10]. Identifying which antigens evoke one or more of these immune pathways therefore remains a key step in developing a subunit-based CBPP vaccine [11]. Phage display [12] makes it possible to identify antigenic proteins by using antibodies from an immune source to select binding peptides from a large repertoire of random amino acid sequences [13]. Fragmented-genome or “”shotgun”" display libraries [14] can directly identify genes that code for the proteins of which the immunoselected peptides form a part.