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Biomedical subjects

H Schaller

Publications and source records attributed to H Schaller.

At least 19 recordsLinked to original sources

Hepatitis B virus genes and their expression in E. coli.

A composite DNA sequence of regions of hepatitis B virus, determined from a series of recombinant plasmids, reveals the genes for the surface antigen and the core antigen of the virus. The sequence of the core antigen shows it to be a DNA binding protein. The core antigen gene is expressed in Escherichia coli and when injected into rabbits the bacterial product induces antibodies which react with core antigen isolated from human sources.

Amino Acids

Nucleotide sequence of the recognition site of the B-specific restriction modification system in E. coli.

Two sB mutations in the genome of bacteriophage fd were located by sequence analysis in the fd sequence at positions 971 and 6341. Base changes at or close to these positions in phage M13 and in phage fl am 124 also correlate with a loss of sensitivity to B restriction. From the sequence homology between the sequences at the two sB sites the recognition signal for the E. coli B restriction/modification enzzyme is predicted to be: 5' TGA---8N---TGCT 3' 3' ACT---8N---ACGA 5'.

Base Sequence

Nucleotide sequence of the origin of replication of the Escherichia coli K-12 chromosome.

The origin of replication, oriC, of the Escherichia coli chromosome was mapped within a DNA segment of 422 base pairs. The nucleotide sequence of this segment was determined. The source of DNA for the sequence analysis was a minichromosome constructed in vivo, consisting exclusively of chromosomal DNA and a minichromosome constructed by cloning in vitro. The nucleotide sequence of the replication origin is characterized by a high degree of repetitiveness due to both inverted and direct repeats. Sequence homologies were found between portions of the replication origins of E. coli and phages lambda and G4. This suggests similarities in some steps in the initiation of replication of the different replicons.

Base Sequence

Transposition of a DNA sequence determining kanamycin resistance into the single-stranded genome of bacteriophage fd.

Derivatives of bacteriophages fd which transduce kanamycin resistance were selected after growth of the phage in an E. coli strain that carried transpoon 5 (Tn5). Different clones of transducing phage and their DNAs were characterized by gel electrophoresis, electron microscopy, and by their ability to multiply in the absence of helper phage. Integration of the intact transposon into the full size phage genome was correlated with an increase in size of the phage particle from 0.95 mu to 1.7 mu, and with the appearance in the phage DNA of the stem loop structure characteristic for single-stranded Tn5 DNA. In non-defective phages the site of insertion was mapped by heteroduplex analysis within the intergenic region of the phage genome. Defective transducing phages were characterized as an insertion of Tn5 into a phage gene, and/or as a partial deletion or duplication of phage and transposon DNA. The size of the transducing phage from different defective clones varied from 0.6 mu to 3.0 mu and was directly proportional to the DNA content. These results demonstrate that filamentous bacteriophage are highly capable to replicate and package very different amounts of foreign DNA.

Coliphages

Structure of the orgin of DNA replication of bacteriophage fd.

An RNA-polymerase-protected DNA fragment of 125 nucleotides from the origin of single-strand to double-strand replication of bacteriophage fd (ori-DNA) was located on the physical map of the phage genome. A stretch of 187 base pairs of DNA including the ori-DNA was sequenced. This DNA segment contains regions with a highly asymmetric pyrimidine/purine distribution next to regions with 2-fold symmetry that form stable hairpin structures in the viral DNA strand.

Base Sequence

An RNA transcribed from DNA at the origin of phage fd single strand to replicative form conversion.

Phage fd DNA complexed with DNA binding protein I was used by Escherichia coli RNA polymerase (nucleoside triphosphate:RNA nucleotidyltransferase, EC 2.7.7.6) to synthesize an RNA at the origin of single strand to double strand replication. The isolated ori-RNA gave a simple fingerprint after nucleolytic digestion and has a length of about 30 nucleotides. The characterization of the oligonucleotides from the nuclease digest and the extension of the ori-RNA with DNA polymerase I and subsequent restriction of the DNA gave its exact localization in the fd genome, and its total sequence was deduced from the known DNA sequence in this region.

Bacteriophages

Nucleotide sequence of bacteriophage fd DNA.

The sequence of the 6,408 nucleotides of bacteriophage fd DNA has been determined. This allows to deduce the exact organisation of the filamentous phage genome and provides easy access to DNA segments of known structure and function.

Base Sequence

Influence of Mg2+-ions on the properties of rat heart mitochondria in dependence on the preparation.

1. The preparation of rat heart mitochondria with Potter-Elvehjem homogenizer results in mitochondria showing stimualtion of respiration induced by Mg2+. This stimualtion is neither caused by adherent hexokinase nor by energy-dependent magnesium accumulation (Mg2+ content in the presence of 10 mM glutamate: 22 nmoles/mg protein; in the presence of glutamate plus antimycin A 21 nmoles/mg protein). 2. The effect of added magnesium is excluded by addition of carboxyatractyloside. This demonstrates the activity of an ATPase outside of the mitochondrial inner membrane. 3. A simple and rapid method for the preparation of Mg2+-insensitive rat heart mitochondria is presented. The minced heart is pressed through a normal syringe and then treated with trypsin. 4. A comparison of mitochondria of both preparations shows that there is no difference in magnesium content and no energy-dependent magnesium influx.

Adenylate Kinase

The dependence on the extramitochondrial ATP/ADP-ratio of the oxidative phosphorylation in mitochondria isolated by a new procedure from rat skeletal muscle.

A simple method for preparation of rat skeletal muscle mitochondria is presented using gentle mechanical homogenization in a syringe and nagarse treatment (EC 3.4.4.16). This method enables the preparation of skeletal muscle mitochondria, whose outer membrane is intact to 95%. Furthermore, with mitochondria prepared by this method the regulation of respiration and phosphorylation by the extramitochondrial ATP/ADP-ratio can be demonstrated. In accordance to rat liver and heart mitochondria and to mitochondria of rabbit reticulocytes, the regulation by the extramitochondrial ATP/ADP-ratio lies in the range from 5 (corresponding to 98% of the maximum respiration) to 100 (corresponding to state 4). At extramitochondrial ATP/ADP-ratios from 0.01 to 1 the respiration rate is nearly constant (maximum rate of respiration).

Adenosine Diphosphate

Interaction of RNA polymerase with promoters from bacteriophage fd.

Replicative form DNA of bacteriophage fd, which had been fragmented with the restriction endonuclease II from Hemophilus parainfluenzae (endo R- HpaII), was reacted with Escherichia coli RNA polymerase; the resulting stable preinitiation complexes were analysed using the filter binding assay followed by gel electrophoresis. At 120mM KCL the first-order rate constants for complex decay were determined to be 10(-2)-10(-6)s-1. The second-order rate constants for complex formation were found to be about 10(6) -10(7) M-1 s-1. From these values association constants for the individual promoters were calculated to be 2 x 10(-8) -2 x 10(-11) M-1. The rate of formation and the stability of promoter complexes was enhanced in superhelical DNA. No evidence was found for stable promoter-specific closed complexes consisting of enzyme and helical DNA. This and the kinetic data suggest that the unwinding of base pairs is already important early in promoter selection, and not only for the formation of the final open complex. The initiation of RNA synthesis form the preinitiation complex was faster than complex dissociation and complex formation for all promoters. Consequently, the initiation efficiency of a promoter is determined by the rate of complex formation, and not by its 'affinity' for the enzyme. No correlation was found between the relative order of the fd promoters for the binding and the dissociation reaction. This is explained by different structural determinants, for the two reactions, which are located in different parts of the promoter DNA.

Base Sequence

[Pre-operative embolisation of vascular tumours in the head (author's transl)].

Trans-vascular, pre-operative embolisation in order to reduce blood flow was carried out with excellent results in a patient with a naso-pharyngeal fibroma, and with relatively good results in a patient with a pterion meningioma. This technique can be carried out following the diagnostic angiogram through the same catheter; it reduces operative blood loss, improves haemostasis and the surgeon's view of the operative field. The method can be used simply and without risk in any case where the tumour blood supply is derived exclusively or predominantly from branches of external carotid artery.

Adult

[The aetiology and management of the frontal sinus mucopyocoele (author's transl)].

The pathogenesis of frontal sinus mucopyocoele is assessed in 33 patients seen between 1958 to 1972. In 45% there has been previous frontal sinus surgery. The importance of the frontonasal duct and healthy mucosa is stressed. The unexpectedly high number of mucopyocoeles following surgery permitted critical assessment of the procedures employed. The most successful and least complicated of which involved preservation or reconstruction of a drainage canal, removal of the obstructing middle turbinate and use of an indwelling drainage tube for at least 6 months. Regular follow-up and adequate early treatment of intercurrent infection are essential.

Frontal Sinus

Nucleotide sequence of an RNA polymerase binding site from the DNA of bacteriophage fd.

The primary structure of a strong RNA polymerase binding site in the replicative form DNA of phage fd has been determined by direct DNA sequencing. It is: (see article). The molecule contains regions with 2-fold symmetry and sequence homologies to promoter regions from other DNAs. The startpoint of transcription is located in the center of the binding site.

Base Sequence