During the epidemic outbreak in the region of Greifswald in the winter 1974/75, we found influenza virus variants which showed differences in the electrophoretic mobility of HA. Among the 25 isolates 13 were of slower and 12 of higher mobility. HA1 of 6 isolates was studied by determining the number of the carbohydrate side chains and by direct sequencing of vRNA. Evidence is presented that variants showing a slower electrophoretic mobility of HA1 had consistently acquired a seventh carbohydrate side chain at Asn 126 in epitope A. All the isolates differed from the reference strain A/Port Chalmers/1/73 by the loss of the oligosaccharide at Asn 81. The field strain A/Dresden/3/71 possessed only 5 oligosaccharides in HA1. These results suggest that changes in glycosylation are an important mechanism in the structural variation underlying antigenic drift of HA.
Summary:In this paper we report that a combination of noncontact and contact atomic force microscopy is a convenient and reliable method for imaging and dissecting single plasmid deoxyribonucleic acid (DNA) strands on mica at ambient conditions without leaving feedback and without damage to the scanning tips. The width and thickness measured at different points of the DNA strands agree with literature data and are the same before and after dissection.
Electrophoretic mobility differences in polyacrylamide gels were detected between (35S)-methionine-labelled nucleoproteins (NPs) induced in monolayer cells by 15 human and 4 avian reference strains of influenza viruses. The (35S)-methionine-labelled tryptic peptides of nucleoproteins of these strains were also analyzed by peptide mapping technique. Based on several detectable hydrophilic peptides the NPs could be arranged in 7 clearly differentiable groups. After radioiodination of NPs from 4 human and 3 avian reference strains the tryptic peptide patterns showed one clear difference between human and avian strains.
Summary: Near-contact mode atomic force microscopy (AFM) imaging leads to sharper representations of DNA double strands on mica imaged at ambient conditions compared with noncontact mode AFM. Phase shift was used for feedback control yielding height information using a simple model calculation. No contact between tip and sample occurs. Measured DNA widths were up to four times smaller than measured with the same AFM tip in noncontact mode at ambient condition.
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