[en] The dhlIVa gene coding for DehIVa was expressed in Escherichia coli and the protein was purified and crystallized using the hanging-drop method

[en] The ochre mutant oc9 of bacteriophage PHI X174 was irradiated with γ-rays and the revertants were assayed on unirradiated and UV-irradiated host bacteria carrying an amber suppressor. The yield of revertants (amber + wild type) was higher on UV-irradiated than on unirradiated bacteria, showing that γ-irradiated PHI X174 was subjected to W-mutagenesis. For oc9 γ-irradiated in the presence of oxygen the fraction of amber mutants among the revertants was lower when mutants were scored on UV-irradiated bacteria than when assayed on unirradiated indicator cells. The same fraction of ambers was obtained when mutants were assayed on unirradiated and UV-irradiated samples of a recA indicator strain. UV-irradiated PHI X174 showed a similar phenomenon. These results suggest that the specificity with regard to insertion of bases opposite radiation damage in PHI X174 DNA is different for host cells in which SOS repair has been induced and cellls in which SOS repair is not operative. (orig.)

[en] Some commercially important vinyl derivatives are produced by the decarboxylation of phenolic acids. Enzymatically, this process can be achieved by phenolic acid decarboxylases (PADs), which are able to act on phenolic acid substrates such as ferulic and p-coumaric acid. Although many microbial PADs have been characterized, little is known regarding their plant homologs. Transcriptome sequencing in the liverworts has identified seven putative PADs, which share a measure of sequence identity with microbial PADs, but are typically much longer proteins. Here, a PAD-encoding gene was isolated from the liverwort species Conocephalum japonicum. The 1197 nt CjPAD cDNA sequence was predicted to be translated into a 398 residue protein. When the gene was heterologously expressed in Escherichia coli, its product exhibited a high level of PAD activity when provided with either p-coumaric or ferulic acid as substrate, along with the conversion of caffeic acid and sinapic acid to their corresponding decarboxylated products. Both N- and C-terminal truncation derivatives were non-functional. The transient expression in tobacco of a GFP/CjPAD fusion gene demonstrated that the CjPAD protein is expressed in the cytoplasm. It is first time a PAD was characterized from plants and the present investigation provided a candidate gene for catalyzing the formation of volatile phenols.

[en] Fresh, chilled chicken carcasses were irradiated at 2.5 kGy and stored at 40 degree C. At intervals samples were withdrawn for microbial, chemical and sensory evaluation. Result showed that combination of a 2.5 kGy irradiation dose and storage at 4 degree C were adequate for a radicidised chicken process. Immediately after irradiation, the microbial spoilage was reduced by at least 4 log cycles. The carcasses were qf excellent quality for at least 16 days of storage and were free from Salmonella and other food pathogens. Changes in chemical composition (moisture, fat, protein, ash and amino acids) and sensory quality of chicken carcasses irradiated at 2.5 kGy were not significant. Therefore the dose of 2.5 kGy should be the target for chilled chicken irradiation process

No abstract available

[en] Mlc, a transcriptional regulator from E. coli, has been expressed, purified and crystallized. Diffraction data were collected to 2.9 Å from an Se-derivative crystal

[en] We describe a general and simple modification to the standard M9 minimal medium recipe that leads to an approximate twofold increase in the yield of heterologously expressed proteins in Escherichia coli BL21(DE3) bacteria. We monitored the growth of bacteria transformed with plasmids for three different test proteins in five minimal media with different concentrations of buffering salts and/or initial media pH. After purification of the over-expressed proteins, we found a clear correlation between the protein yield and change in media pH over time, where the minimal media that were the most buffered and therefore most resistant to change in pH produced the most protein. And in all three test protein cases, the difference in yield was nearly twofold between the best and worst buffering media. Thus, we propose that increasing the buffering capacity of M9 minimal media will generally lead to a similar increase for most of the proteins currently produced by this standard protein expression protocol. Moreover, we have qualitatively found that this effect also extends to deuterated M9 minimal media growths, which could lead to significant cost savings in these preparations.

[en] The overproduction, purification, crystallization and preliminary crystallographic studies of the native and selenomethionine-labelled P1 domain are reported here as a first step towards the elucidation of the molecular mechanism of YidC as a membrane-protein insertase. In Escherichia coli, the biogenesis of inner membrane proteins (IMPs) requires targeting and insertion factors such as the signal recognition particle (SRP) and the Sec translocon. Recent studies have identified YidC as a novel and essential component involved in membrane insertion of IMPs both in conjunction with the Sec translocon and as a separate entity. E. coli YidC is a member of the YidC (in bacteria)/Oxa1 (in mitochondria)/Alb3 (in chloroplasts) protein family and contains six transmembrane segments and a very large periplasmic domain P1. The overproduction, purification, crystallization and preliminary crystallographic studies of the native and selenomethionine-labelled P1 domain are reported here as a first step towards the elucidation of the molecular mechanism of YidC as a membrane-protein insertase

[en] Abstract only

[en] Several crystals of the Grb2-like C-terminal SH3 domain in complex with a motif peptide from the SLP-76 protein were obtained and characterized. The Grb2-like adaptor protein GADS is composed of an N-terminal SH3 domain, an SH2 domain, a proline-rich region and a C-terminal SH3 domain. GADS interacts through its C-terminal SH3 domain with the adaptor protein SLP-76, thus recruiting this protein and other associated molecules to the linker for activation of T-cell (LAT) protein. The DNA encoding the C-terminal SH3 domain of GADS (GADS-cSH3) was assembled synthetically using a recursive PCR technique and the protein was overexpressed in Escherichia coli, refolded and purified. Several crystals of this domain in complex with the SLP-76 peptide were obtained and characterized