We have developed a primer set for a prokaryotic differential display of mRNA in the Enterobacteriaceae group. Each combination of ten 10mer and ten 11mer primers generates up to 85 bands from total Escherichia coli RNA, thus covering expressed sequences of a complete bacterial genome. Due to the lack of polyadenylation in prokaryotic RNA the type T11VN anchored oligonucleotides for the reverse transcriptase reaction had to be replaced with respect to the original method described by Liang and Pardee [ Science , 257, 967-971 (1992)]. Therefore, the sequences of both the 10mer and the new 11mer oligonucleotides were determined by a statistical evaluation of species-specific coding regions extracted from the EMBL database. The 11mer primers used for reverse transcription were selected for localization in the 3'-region of the bacterial RNA. The 10mer primers preferentially bind to the 5'-end of the RNA. None of the primers show homology to rRNA or other abundant small RNA species. Randomly sampled cDNA bands were checked for their bacterial origin either by re-amplification, cloning and sequencing or by re-amplification and direct sequencing with 10mer and 11mer primers after asymmetric PCR.
CMV-DNA in smooth muscle cells induces local growth factor expression as well as endothelial activation, both of which can promote the progression of atherosclerosis. Since traditional atherogenic risk factors increase the likelihood of aortic CMV manifestation, we suggest that CMV plays a crucial role in mediating the progression of atherosclerosis.
1-Deoxy-3,4:5,6-di-O-isopropylidene-D-arabino-hex-2-ulose reacted with N,N-dimethylformamide dimethyl acetal and tert-butoxy[bis(dimethylamino)]methane, respectively, to furnish (E)-1,2-dideoxy-1-dimethylamino-4,5:6,7-di-O-isopropylidene-Darabino-hept-1-en-3-ulose. This push-pull activated heptenulose underwent ring closure reactions with various N,N'-nucleophiles like hydrazine hydrate, amidinium, guanidinium and isothiouronium salts in the presence of bases to yield pyrazole and pyrimidine derivatives, respectively, all of which derivatized with a di-O-isopropylidenated tetritol side chain. The isopropylidene groups were removed with y
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