Biomedical subjects

R Cannio

Publications and source records attributed to R Cannio.

22 records · Page 2Linked to original sources

PubMed1994

A few amino acid substitutions are responsible for the higher thermostability of a novel NAD(+)-dependent bacillar alcohol dehydrogenase.

The gene adh-hT encoding a thermostable and thermophilic NAD(+)-dependent alcohol dehydrogenase (ADH) from the novel and more thermophilic Bacillus stearothermophilus LLD-R strain was cloned and its nucleotide sequence determined. The deduced protein sequence shows remarkable amino acid substitutions when compared to the sequence of the protein isolated from strain NCA1503 and significant similarity with the highly thermostable ADH from the thermoacidophilic archaebacterium Sulfolobus solfataricus. The alignment of these sequences led to the identification of three amino acid replacements probably responsible for the higher thermostability of the novel bacillar ADH. Adh-hT gene expression in Escherichia coli, a fast purification procedure and the characterization of the recombinant enzyme are also described.

Alcohol Dehydrogenase↗

PubMed1994

Gene expression of a thermostable beta-galactosidase in mammalian cells and its application in assays of eukaryotic promoter activity.

The gene (lacS) encoding a thermostable beta-galactosidase enzyme (Ss beta-gal) from the archaebacterium Sulfolobus solfataricus has been cloned and expressed in simian CV1 and murine NIH3T3 cell lines. The recombinant protein is an active enzyme that shows the same properties of thermophilicity and thermostability as the wild-type Ss beta-gal and has no cytotoxic effect on the host cells. Its possible use as a reporter gene is also proposed, and a comparison with other reporter gene systems is made.

3T3 Cells↗

PubMed1993

Stability and activity of a phenol oxidase from the ligninolytic fungus Pleurotus ostreatus.

Three different phenol oxidases produced by the basidiomycete fungus Pleurotus ostreatus have been isolated and their main structural, enzymatic and physico-chemical properties characterized. Studies have focused on the most abundantly secreted of these proteins, a copper-enzyme specific towards ortho-diphenol substrates. This protein was purified to homogeneity and part of its primary structure determined by direct protein sequencing. The influence of pH, temperature and presence of water-soluble or water-insoluble organic solvents on the activity and stability of the enzyme were also investigated. These data can be used for applying bioreactors to problems of environmental concern such as waste-water treatment.

Amino Acid Sequence↗

PubMed1991

A cell-type specific and enhancer-dependent silencer in the regulation of the expression of the human urokinase plasminogen activator gene.

A transcriptional silencer has been identified in the 5' regulatory region of the human urokinase plasminogen activator (uPA) gene. This region is able to block transcription from the human u-PA as well as the rabbit beta-globin promoters in a cell type specific and orientation independent way. The silencer is enhancer dependent and is active in two cell lines (HeLa and CV-1) which produce little if any uPA, but not in the high uPA producer PC3. Silencing activity and enhancer dependence can be separated: the silencing activity has been localized to the DNA fragment -660 to -536, while the enhancer dependence is located in the -536 to -308 fragment. The DNA sequence of the silencer region contains an element that closely resembles the TGF-beta responsive negative element TIE.

Animals↗