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M Dalrymple

Publications and source records attributed to M Dalrymple.

7 recordsLinked to original sources
PubMed1998

Production of biologically active salmon calcitonin in the milk of transgenic rabbits.

Salmon calcitonin (sCT) is an example of one of the many bioactive peptides that require amidation of the carboxy terminus for full potency. We describe a method for the production of amidated sCT in the mammary gland of transgenic rabbits. Expression of a fusion protein comprising human alpha lactalbumin joined by an enterokinase cleavable linker to sCT was directed to the mammary gland under the control of the ovine beta lactoglobulin promoter. C-terminal amidation in vivo was achieved by extending the sCT by a single glycine residue that provides a substrate for endogenous amidating activity in the mammary gland. Full characterization of the released sCT demonstrated it to be equivalent to synthetic standard in terms of structure, purity, and potency.

Amides↗
PubMed1996

High-level expression of recombinant human fibrinogen in the milk of transgenic mice.

Fibrinogen is a complex plasma protein composed of two each of three different polypeptide chains. We have targeted expression of r-human fibrinogen to the mammary gland of transgenic mice. Three expression cassettes, each containing the genomic sequence for one of the three human fibrinogen chains controlled by sheep whey protein beta-lactoglobulin promoter sequences, were coinjected into fertile mouse eggs. Southern blot analysis demonstrated that more than 80% of the transgenic founders contained all three fibrinogen genes. Reducing sodium dodecyl sulfate polyacrylamide gel electrophoresis of milk from the highest producing founder animal demonstrated the presence of human fibrinogen subunits at concentrations of 2000 micrograms/ml. In several animals with a balanced ratio of the individual fibrinogen subunits, up to 100% of the protein was incorporated into fully assembled fibrinogen hexamers. Incubation of the transgenic milk with thrombin and factor XIII resulted in a cross-linked fibrin clot, indicating that a major portion of the secreted fibrinogen was functional. These studies represent the first report of high-level biosynthesis and secretion of a functional, complex, hexameric protein in the milk of a transgenic animal.

Animals↗
PubMed1993

Maker's mark.

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Base Sequence↗
PubMed1993

Model for assessing the risk of introducing brucellosis into a brucellosis-free area.

A risk assessment model is presented, for use by local decision-makers to aid the evaluation of proposed changes in existing brucellosis eradication or control programmes. This model provides a format and structure for gathering and analysing data. The model uses data which are generally available and accessible, so that minimum time, expense and effort are required for collection. The use of this model enables an estimation of the risk of introduction of brucellosis into a non-infected population, based on the probability of importing the agent and subsequent spread, given the existence of specified surveillance and control measures. The model creates a point estimate of the risk associated with a given set of conditions.

Animals↗
PubMed1992

Expression of human alpha 1 antitrypsin in transgenic sheep.

We have recently described the production of large amounts (< or = 65 grams per litre) of enzymatically active human alpha 1 antitrypsin in the milk of transgenic sheep (Wright et al., 1991). Here, we describe in more detail the expression of the human protein in the milk of these animals throughout the lactation period. Human alpha 1 antitrypsin is also found at much lower levels in the plasma of transgenic ewes before, during and after lactation. It is also detected in male plasma at very low levels. We have previously shown human alpha 1 antitrypsin purified from transgenic sheep milk to be indistinguishable from commercially available human plasma derived alpha 1 antitrypsin in terms of gross sugar content and in vitro activity. Here we extend this comparison to more detailed analyses of glycosylation state, amino-terminal sequence, pI value, and molecular weight determination by mass spectrometry.

Amino Acid Sequence↗
PubMed1985

Reservoir animals for nephropathia epidemica in Norway: indications of a major role for the bank vole (C. glareolus) in comparison with the woodmouse (A. sylvaticus).

Small rodents were collected live in two different locations within a nephropathia epidemica (NE) endemic area, and tested for both antiviral serum antibodies and viral antigens in lung sections. In one location, only Apodemus sylvaticus (woodmice) were found in the traps, in the other, both A. sylvaticus and Clethrionomys glareolus (bank voles) were collected. Among the woodmice from the former location the prevalence of NE virus markers was significantly lower than for either woodmice or bank voles from the other location, and no NE antigen-positive animals was found. The woodmice co-existing with bank voles had a lower prevalence of NE antigen and antibodies than the bank voles, and fewer woodmice had both antibodies and antigen. The results emphasize the important role of bank voles as a major NE virus reservoir and probable source of human infections.

Animals↗