Biomedical subjects

G B Poulsen

Publications and source records attributed to G B Poulsen.

5 recordsLinked to original sources

PubMed1993

A useful cloning vector for Bacillus subtilis.

We have constructed plasmid pDN1050, a new small cloning vector for Bacillus subtilis. pDN1050 harbors the origin of replication of Staphylococcus aureus plasmid pUB110 and the chloramphenicol resistance gene of S. aureus plasmid pC194. The plasmid is segregationally and structurally stable. Plasmid pDN1370, a low copy number mutant of pDN1050 was isolated and shown to harbor a mutation in the repA gene of the replication protein.

Bacillus subtilis↗

PubMed1992

The 5' untranslated region from pea seedborne mosaic potyvirus RNA as a translational enhancer in pea and tobacco protoplasts.

We have exploited the transient expression of foreign genes introduced into plant protoplasts to investigate the effect of the pea seedborne mosaic potyvirus (PSbMV) 5' untranslated region (5'UTR) on the level of gene expression in pea and tobacco protoplasts. The plant viral 5'UTRs were found to increase translation significantly in comparison to a plasmid containing no 5'UTR of viral origin. The enhancement effect of the 5'UTRs of PSbMV and tobacco etch potyvirus (TEV) was found to be similar in pea and tobacco protoplasts, indicating a host-independent role of the potyviral 5'UTRs in enhancing gene expression. Translational enhancement of the two potyviral 5'UTRs was similar to that of the 5'UTR of tobacco mosaic virus (TMV). This observation makes it attractive to use potyviral 5'UTRs as general translational enhancers in future genetic transformations of plants.

Base Sequence↗

PubMed1991

Cloning of a chromosomal alpha-amylase gene from Bacillus stearothermophilus.

We have cloned and sequenced a gene for a heat-stable alpha-amylase from a natural isolate of Bacillus stearothermophilus. Previously, it had been shown that B. stearothermophilus amylase genes may be harboured on indigenous plasmids. We have found that our isolate harbours the amylase gene only on the chromosome and not on its indigenous plasmid.

Amino Acid Sequence↗

PubMed1990

In vivo genetic engineering: homologous recombination as a tool for plasmid construction.

This paper describes a novel method for creating exact DNA fusions between any two points in a plasmid carried in Bacillus subtilis. It exploits the homologous in vivo recombination between directly repeated sequences that can be established by insertion of a synthetic oligodeoxyribonucleotide. The method was used to enhance the productivity in B. subtilis of a cloned alpha-amylase (Amy)-encoding gene originating from Bacillus stearothermophilus. Thus, an exact fusion between nucleotide sequences encoding the expression signals, including the signal peptide, of a Bacillus licheniformis Amy-encoding gene and the mature Amy of B. stearothermophilus, was created. The resulting hybrid translational product was processed correctly in B. subtilis during secretion, giving rise to an Amy identical to the mature Amy secreted by B. stearothermophilus.

Amino Acid Sequence↗

PubMedDate not supplied

Cloning and expression of an amylase gene from Bacillus stearothermophilus.

In the industrial process of liquefying starch to make glucose or high fructose syrups, it is crucial that the amylase used is stable and active at about 105 degrees C at pH 6.5 or preferentially at a lower pH. The amylase from Bacillus licheniformis is well suited for this purpose but it is possible that other amylases might perform even better. Therefore, we cloned and characterized amyS encoding a heat-stable alpha-amylase from Bacillus stearothermophilus. Using a newly developed method for creating exact gene fusions by in vivo recombination, we attempted to increase expression of amyS in Bacillus subtilis. However, only by introducing the amyS gene into B. licheniformis, we obtained significantly better yields.

Bacillus↗