[History of the development of endoscopy].
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Biomedical subjects
Publications and source records attributed to V Paces.
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The 12,200-bp sequence of the late region of bacteriophage PZA was determined. Open reading frames (ORFs) and potential ribosome-binding sites were found in this region and the ORFs were assigned to eleven late genes. A potential bidirectional transcriptional terminator was found and its possible function is discussed.
We have sequenced the rightmost 2079 bp of the Bacillus subtilis phage PZA genome. This region encompasses the right early region. We compared it with the homologous region of phage phi 29. Six open reading frames (ORFs) were found in this region of PZA and one of them was assigned to gene 17. Analysis of putative ribosome-binding sites and comparison with phi 29 ORFs indicate that at least some of the remaining ORFs could encode proteins. Corresponding genes were not identified so far by genetic methods. Promoter candidates in the right early region of PZA were found and compared to phi 29 promoters. The sequenced region together with previously determined sequences [Paces et al., Gene 38 (1985) 45-56 and 44 (1986) 107-114] completes the entire 19,366-bp sequence of phage PZA genome.
The 12,177-bp nucleotide (nt) sequence of the late region of Bacillus phage luminal diameter 29 genome was determined. This sequence completes the entire 19,285-bp sequence of phage luminal diameter 29 DNA. Eleven open reading frames were found in this region, and these were assigned to eleven late genes. Ribosome-binding sites and a potential transcriptional promoter and terminator are considered. The nt sequence was compared to the homologous region of the closely related phage PZA and tolerated variations at the nt and amino acid (aa) level were evaluated. The most frequent changes are silent nt substitutions in the third position of codons, but aa substitutions are also found.
The DNAs of phages phi 29, phi 15 and PZA of Bacillus subtilis were analysed with restriction enzymes EcoRI, HpaI and HindIII. A method was used which permits parallel physical mapping of all three phages, from both ends of their linear genomes. The method is based on transfer of partially digested DNA to DBM paper and sequential hybridization with labelled terminal fragments. It follows from the comparison of the physical maps that phages phi 29, phi 15 and PZA are closely related and that they probably have arisen from a common ancestor by accumulation of point mutations.
The 5200-bp nucleotide sequence of the major early region of bacteriophage PZA has been determined. Open reading frames (ORFs) and potential transcriptional and translational regulatory signals were found in this region. The sequence was compared with the known sequence of the homologous region of the closely related phage phi 29 (Yoshikawa and Ito, 1982). This comparison permitted a more accurate assignment of several ORFs and regulatory signals.
Enkephalins are pentapeptides with opioid activity that have been found in brain and other neural tissues. They are released by proteolytic processing of the proenkephalin, which contains several enkephalin sequences each flanked by pairs of basic amino acid (aa) residues. We have constructed an artificial variant of the proenkephalin gene by concatenation of synthetic oligodeoxynucleotides (oligo) coding for Met-enkephalin preceded by two arginines. One of the resulting structures, containing eleven enkephalin sequences separated by pairs of arginine codons, was cloned in the expression vector pRE31. The biological activity of enkephalin was detected after the digestion of the isolated plasmid-coded protein with trypsin and carboxypeptidase B. The product of the synthetic gene may thus serve as a defined simplified substrate for the study of the not yet fully understood enzymatic mechanisms of proenkephalin processing.
Morphologically identical phages PZA, PZE , phi 29, and phi 15 can be distinguished by the neutralization test with rabbit antisera and by host range specificity. Each member of this phage group contains 18 kb double-stranded linear DNA carrying proteins covalently attached to its 5' ends. Physical maps of their DNA constructed with the use of restriction endonucleases EcoRI, HpaI, HindIII, BspRI, and XbaI and DNA-DNA hybridization experiments show a closer relationship between phi 29 and PZE than between phi 29 and phi 15 or phi 29 and PZA. phi 15 is closer to PZA than to phi 29. This conclusion is supported by analysis of differential denaturation profiles of the phage DNAs. Sequencing of selected parts of phi 29 and PZA DNAs reveals 93% homology with a preference for synonymous base replacements (silent mutations) randomly distributed in the coding regions. Using promoter-probe plasmid pPV33 -H the region functioning as a promoter in E. coli was localized on the smallest EcoRI fragment of PZA and phi 29 DNAs. Comparison of the nucleotide sequence of this region with known promoters of B. subtilis shows extensive homologies with at least two types of promoters of different specificities, namely those recognized by factors sigma 28 of B. subtilis and sigma gp28 of phage SPO1 . These promoter-like regions overlap and the whole sequence is conserved in both phages.
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Cytokinin oxidase has been partially purified from cultured tobacco tissue. This enzyme converts N6-(delta2-isopentenyl)-adenosine to adenosine. The reaction is inhibited by the two isomers of ribosylzeatin [n6-4-hydroxy-3-methylbut-2-enyl)adenosine]. Trans-ribosylzeatin inhibits the reaction more than the cis-isomer.
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