Recombinant Globular Domain of TcpA Pilin from El Tor: Recovery from Inclusion Bodies and Structural Characterization.

The production of recombinant proteins in cells is often hampered by aggregation of newly synthesized proteins and formation of inclusion bodies. Here we propose the use of transverse urea gradient electrophoresis (TUGE) in testing the capability of folding of a recombinant protein from inclusion bodies dissolved in urea. A plasmid encoding the amino acid sequence 55-224 of TcpA pilin (C-terminal globular domain: TcpA-C) from El Tor enlarged by a His-tag on its N-terminus was expressed in cells.

Development and Evaluation of a One-Step Quantitative RT-PCR Assay for Detection of Lassa Virus.

Lassa fever is a severe viral hemorrhagic illness caused by Lassa virus. Based on estimates, the number of LASV infections ranges from 300,000 to 500,000 cases in endemic areas with a fatality rate of 1%. Development of fast and sensitive tools for the control and prevention of Lassa virus infection as well as for clinical diagnostics of Lassa fever are crucial.

Development of a diagnostic test for Yersinia pestis by the polymerase chain reaction.

A 501 bp caf1 gene fragment and a 443 bp of pla gene fragment carried by 100 kb (pFra) and 10 kb (pPst) species-specific extrachromosomal replicons, respectively, were used as targets to study the conditions under which DNA amplification by polymerase chain reaction (PCR) may be applied to detect and identify Yersinia pestis DNA in cell lysates of pure cultures and biological samples. The sensitivity limit of PCR with the crude cell lysates of Y. pestis EV was estimated as 10-50 cfu in reaction mixture.

[In vivo and in vitro study of plasmid fragments from Ca2+-dependent Yersinia pestis (Lehmann, Neumann)].

The bank of the HindIII, EcoRI and PstI fragments of Yersinia pestis Ca(2+)-dependence plasmid (pCaD) was constructed and used as molecular probes, combined with the restriction analysis to map pCaD. Experiments on laboratory animals showed two subfragments of the fifth HindIII fragment to impart virulence, invasiveness to the plasmidless avirulent strains of Y. pestis, Y.