This article describes the construction of a set of versatile expression vectors based on the In-Fusion cloning enzyme and their use for high-throughput cloning and expression screening. Modifications to commonly used vectors rendering them compatible with In-Fusion has produced a ligation-independent cloning system that is (1) insert sequence independent (2) capable of cloning large PCR fragme...

BACKGROUND In the United Kingdom (UK), the National Treatment Agency for Substance Misuse (NTA) considers retention to be the best available measure of drug treatment effectiveness. Accordingly, the NTA has set local treatment systems the annual target of retaining 75% of clients for 12 weeks or more, yet little assessment of this target or factors that improve retention has occurred. This stud...

Brevinin-2R, a member of a new family of antimicrobial peptides isolated from the skin of Rana ridibunda, displays antimicrobial activity against bacteria and fungi. In this study, we have used an assembly PCR method for the fast and extremely accurate synthesis of the brevinin-2R gene. A total of six primers were assembled in a single step PCR, and the assembly was then amplified by PCR to pro...

Titania nanotube arrays (NTA) generated from anodizing processes are tested as the substrate for surface-assisted laser desorption/ionization mass spectrometry (SALDI MS). The background generated from titania NTA is very low, making the approach suitable for the analysis of small molecules. The upper detectable mass is approximately 29 kDa. Homogeneous sample deposition leads to good shot-to-s...

The ftsZ gene from Xanthomonas oryzae pv. Oryzae was amplified by PCR with the specific primers, and the recombinant plasmid pET-22b-ftsZ was constructed successfully. The FtsZ with a 6× His tag was overexpressed in a soluble form in Escherichia coli BL21 and purified through a Ni-NTA agarose column. The purified recombinant FtsZ showed a single band on SDS-PAGE with an apparent molecular mass ...

Silicon nanowire (Si NW)-based field effect transistors (FETs) have shown great potential as biosensors (bioFETs) for ultra-sensitive and label-free detection of biomolecular interactions. Their sensitivity depends not only on the device properties, but also on the function of the biological recognition motif attached to the Si NWs. In this study, we show that SiNWs can be chemically functional...

We study the solvation of amino acids in pure-osmolyte and mixed-osmolyte urea and trimethylamine N-oxide (TMAO) solutions using molecular dynamics simulations. Analysis of Kirkwood-Buff integrals between the solution components provides evidence that in the mixed osmolytic solution, both urea and TMAO are mutually excluded from the amino acid surface, accompanied by an increase in osmolyte-osm...

During infection, poliovirus binds to its receptor and undergoes conformational changes that ultimately result in release of the genomic RNA into the cell. The details of this process are poorly understood. Tuthill et al. (p. 172–180) report the development of a simple model system in which liposomes containing NTA head groups are used to capture His-tagged ectodomains of the poliovirus recepto...