Fragment C is the C-terminal domain of the heavy chain of tetanus toxin that can promote the immune response against the lethal dose of this toxin. Therefore, this portion can be considered as a candidate vaccine against tetanus infection, which occurs by Clostridium tetani. The present study aimed to compare the expression of tetanus toxin fragment C in Escherichia coli  BL21 (DE3) pLysS cells...

This protocol describes the detection of small RNAs (~10-200 nucleotides) by blot hybridization. The RNA samples, denatured in formamide, are separated by denaturing polyacrylamide gel electrophoresis. Because high-percentage polyacrylamide gels are required to separate RNAs in this size range, it is necessary to perform electrophoretic transfer to positively charged nylon membranes. After tran...

Introduction 2 Deoxyribonucleic Acid to Ribonucleic Acid 4 Ribonucleic Acid to Protein 6 Proteome Handling in Classical and Functional Proteomics 6 Protein Purification 6 Cell Culture 8 Other Source of Samples 8 Subpopulation of Cells 10 Subcellular Components 11 Protein Separation by Two-Dimensional Gel Electrophoresis 12 Principles of Two-Dimensional Gel Electrophoretic Separation 13 Detectio...

Clostridial collagenases are essential biotechnological tissue dissociation agents owing to their ability cleave different types of collagen. Standardization collagenase-based protocols has been hampered by impurities in products manufactured from Clostridium histolyticum . To enhance the purification process, we produced recombinant collagenase classes G and H, taking advantage Escherichia col...

METHODS. Gene targeting in embryonic stem cells was used to generate mouse lines in which the aB gene and its protein product were absent. Gene structure and expression were characterized by genomic Southern blot, immunoblot, and Northern blot analyses, and two-dimensional gel electrophoresis. The gene knockout mice were screened for cataract with slit lamp biomicroscopy, and dissected lenses w...

Current methods for Northern blot analysis to determine mRNA abundance in tissues are tedious and utilize toxic chemicals. The requirement for chemicals of high purity, free of ribonucleases, and the lability of RNA, in comparison to DNA, are additional factors preventing wider use of the technique. For Northern blot analysis, RNA is generally denatured with glyoxal (1) or formaldehyde (2) prio...

Rochalimaea quintana was isolated from the blood of a French human immunodeficiency virus-infected patient with bacillary angiomatosis. The isolate showed the typical growth characteristics of Rochalimaea species and was inert when typical biochemical testing was used. The purpose of the present work was to characterize and compare this new isolate with reference strains of R. quintana, Rochali...

Until the recent discovery of pRF in Rickettsia felis, the obligate intracellular bacteria of the genus Rickettsia (Rickettsiales: Rickettsiaceae) were thought not to possess plasmids. We describe pRM, a plasmid from Rickettsia monacensis, which was detected by pulsed-field gel electrophoresis and Southern blot analyses of DNA from two independent R. monacensis populations transformed by transp...

A method was developed for detection of Listeria monocytogenes by polymerase chain reaction amplification followed by agarose gel electrophoresis or dot blot analysis with a 32P-labeled internal probe. The technique identified 95 of 95 L. monocytogenes strains, 0 of 12 Listeria strains of other species, and 0 of 12 non-Listeria strains.

Quantitative and qualitative modifications of the plasma proteins are of particular importance among the systemic manifestations of rheumatoid arthritis. Increase of a2and y-globulins may be found in most patients (Ropes, Perlmann, Kaufman, and Bauer, 1954) in relation to the onset, evolution, and activity of the disease (Bonomo, 1957). The most important pattern consists generally of a predomi...