We developed a rapid method for extraction of DNA from honey bees, Apis mellifera, and from the parasitic bee mite, Varroa destructor. The advantages include fast processing and low toxicity of the substances that are utilized. We used lysis buffer with nonionic detergents to lyse cell walls and proteinase K to digest proteins. We tested whole thorax, thoracic muscle mass, legs, and antennae fr...

Previous studies have indicated that scrapie infection results in the accumulation of a proteinase K-resistant form of an endogenous brain protein generally referred to as prion protein (PrP). The molecular nature of the scrapie-associated modification of PrP accounting for proteinase K resistance is not known. As an approach to understanding the cellular events associated with the PrP modifica...

BACKGROUND Biofilms are organized communities of microorganisms embedded in a self-produced extracellular polymeric matrix (EPM), often with great phylogenetic variety. Bacteria in the subgingival biofilm are key factors that cause periodontal diseases; among these are the Gram-negative bacteria Fusobacterium nucleatum and Porphyromonas gingivalis. The objectives of this study were to character...

UNLABELLED INTRODUCTION The coexistence of different molecular types of classical protease-resistant prion protein in the same individual have been described, however, the simultaneous finding of these with the recently described protease-sensitive variant or variably protease-sensitive prionopathy has, to the best of our knowledge, not yet been reported. CASE PRESENTATION A 74-year-old Ca...

The presence of organic compounds and high amount of secondary metabolites (polysaccharides, phenolic component, etc.) in mosses cause difficulties in DNA extraction that are followed by problems in PCR reactions. In lower plants, various methods have been used for DNA extraction including silica gel and different commercial kits. These methods mostly use hazardous (like phenol or liquid nitrog...

Proteinase K is widely used in tests for the presence of infectious prion protein causing fatal spongiform encephalopathies. To investigate possible interactions between the enzyme and the functionally important N-terminal prion domain, we crystallized mercury-inhibited proteinase K in the presence of the synthetic peptides GGGWGQPH and HGGGW. The octapeptide sequence is identical to that of a ...

Aqualysin I is the alkaline serine protease isolated from an extreme thermophile, Thermus aquaticus YT-1. We analyzed kinetic properties of aqualysin I, using sixteen kinds of chromogenic succinyl-tripeptide p-nitroanilides as substrates. And we compared the substrate specificity of aqualysin I with those of proteinase K, subtilisin BPN', and subtilisin Carlsberg. We found that aqualysin I had ...

Enhanced canine distemper virus (CDV) nucleoprotein RNA was applied to paraffinembedded tissues from dogs with spontaneous CDV infections. In addition to proteinase K, the tissues were autoclaved in various solutions as a pre-treatment (Trilogy, TBS S3006, H-3301, and S1700) and then compared. The in situ hybridization (ISH) intensity was assessed using integrated optical density (IOD) and tiss...