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R Hauptmann

Publications and source records attributed to R Hauptmann.

30 records · Page 2Linked to original sources
PubMed1987

Structure and expression in Escherichia coli of canine interferon-alpha genes.

Using a human interferon-alpha (IFN-alpha) cDNA probe, several recombinant phages containing type I IFN genes were isolated from a canine genomic library. One of these phages contains two complete CaIFN-alpha genes with identical coding sequences, and a second one a slightly different IFN-alpha gene. The IFN-alpha protein sequences contain six cysteine residues as well as two or three potential N-glycosylation sites. Expression of mature CaIFN-alpha 1 in E. coli results in antiviral activity on dog cells. Genomic analysis using an equine IFN-omega probe and DNA sequencing suggests the deletion of IFN-omega genes from canine genome.

Amino Acid Sequence↗
PubMed1986

[Escherichia coli in biotechnology].

Escherichia coli plays a central role in modern biotechnology i.e. gene technology and fermentation. This article summarizes the basic principles of genetic engineering using human type I interferon genes as an example. The second part describes the advantages and the problems encountered using E. coli as a production organism.

DNA, Bacterial↗
PubMed1986

Molecular cloning and expression in Escherichia coli of equine type I interferons.

Using human interferon-alpha 2 (IFN-alpha 2) and IFN-beta DNA to probe an equine genomic library we isolated recombinant phages containing genes for equine interferon-alpha (EqIFN-alpha), interferon-beta (EqIFN-beta), and interferon-omega (EqIFN-omega). Sequence and hybridization analyses of these genes reveal that the equine genome contains gene families of each of these three type I interferon classes. The mature proteins of EqIFN-alpha are 71-77% homologous to human IFN-alpha polypeptides, and, when expressed in E. coli, possess antiviral activity on both equine and human cells. By contrast, EqIFN-beta is only 59% homologous to its human counterpart and shows activity only on equine cells.

Animals↗
PubMed1985

A novel class of human type I interferons.

The screening of a cDNA library prepared from mRNA of Sendai virus induced Namalwa (human Burkitt's lymphoma) cells, using a human IFN-alpha 2 DNA probe under conditions of low stringency, identified two weakly hybridizing clones containing sequences related to, but discernably different from those of the IFN-alpha class. Sequence and hybridization analysis of these cDNAs as well as expression in E. coli provided evidence that they encode proteins which have the characteristics of IFN type I but which are sufficiently diverged in sequence from both IFN-alpha s and IFN-beta to suggest that they are representatives of a new and distinct class of interferons named interferon-omega. Hybridization of these sequences to genomic DNA reveals that this class contains at least four members.

Base Sequence↗
PubMed1983

Nucleic acid sequence of the region of the genome encoding capsid protein VP1 of neurovirulent and attenuated type 3 polioviruses.

Three closely related strains of poliovirus type 3 have been used to study the molecular basis of attenuation in the currently used Sabin vaccine of this serotype. Plaque-purified derivatives of these strains possess closely similar serological and biochemical properties yet differ markedly in neurovirulence for monkeys. Molecular cloning via an RNA . cDNA method has facilitated comparative nucleotide sequencing. Initial efforts have concentrated on the region of the genome encoding VP1. Only minor structural differences between neurovirulent and attenuated type 3 strains were detected, in contrast to the major differences observed between the vaccine strains of poliovirus type 1 and its virulent precursor P1/Mahoney. These observations suggest that the molecular basis of attenuation of type 3 Sabin vaccine virus does not involve the VP1 polypeptide and, therefore, that mutations conferring the attenuated phenotype probably lie elsewhere in the genome.

Base Sequence↗
PubMed1983

The nucleotide sequence of poliovirus type 3 leon 12 a1b: comparison with poliovirus type 1.

The complete nucleotide sequence of the genome of the Sabin vaccine strain of poliovirus type 3 (P3/Leon 12 a1 b) has been determined from cDNA cloned in E. coli. The genome comprises a 5' non-coding region of 742 nucleotides, a large open reading frame of 6618 nucleotides (89% of the sequence) and a 3' non-coding region of 72 nucleotides. There is 77.4% base-sequence homology and 89.6% predicted amino-acid homology between types 1 and 3. Conservation of all glutamine-glycine and tyrosine-glycine cleavage sites suggests a mechanism of polyprotein processing similar to that established for poliovirus type 1.

Amino Acid Sequence↗
PubMed1983

Poliovirus type 3: molecular cloning of the genome and nucleotide sequence of the region encoding the protease and polymerase proteins.

Overlapping cDNA clones representing the entire genome of poliovirus type 3 have been prepared in E. coli by two separate methods. Cloning of RNA . cDNA hybrids produced a more comprehensive set of clones with generally larger cDNA inserts than cloning of double - stranded cDNA. A restriction map of the entire genome and the nucleotide sequence of 2003 bases from the 3' terminus, comprising the region encoding the protease and polymerase proteins, are presented.

Amino Acid Sequence↗
PubMed1983

Location and primary structure of a major antigenic site for poliovirus neutralization.

We have determined a major antigenic site for virus neutralization on the capsid protein VP1 of poliovirus type 3. Antigenic mutant viruses selected for resistance to individual monoclonal antibodies had point mutations concentrated in a region 277-294 bases downstream from the start of the region of viral RNA coding for VP1. These findings provide the basis for an improved understanding of the molecular basis of virus neutralization.

Amino Acid Sequence↗
PubMedDate not supplied

Nucleotide sequence from neurovirulent and attenuated strains of type 3 poliovirus.

As part of an investigation into the molecular basis of attenuation in the Sabin poliovirus vaccines, the genomes of three strains of type 3 poliovirus differing in neurovirulence were cloned in Escherichia coli. The nucleotide sequence of the region of the genome encoding the virion protein 1 (VP1) polypeptide from each of these strains is presented. Very few changes were observed, a finding suggesting that the mutation(s) involved in the attenuation of type 3 poliovirus probably lies elsewhere in the genome. The complete nucleotide sequence of one of the strains, P3/Sabin, has been obtained and is compared with the published sequences of type 1 poliovirus. Between serotypes 1 and 3, there is 77% homology in nucleotide sequence and 90% homology in the amino acid sequence predicted for viral proteins.

Amino Acid Sequence↗