Biotec Pharmacon Asa

Universitetet I TromsØ—norges Arktiske Universitet

Arcticzymes As

Olav Lanes

Marit Sjo Lorentzen

Morten Elde

Terese Solstad

Yvonne Piotrowski

Atle Noralf Larsen

Nils Peder Willassen

Dag Rune Gjellesvik

Elisabeth Lill Andreassen

Bjørn Altermark

Linda Havdalen

Ingar Leiros

Ronny Helland

Nils Peder Willasen

Per Henrik Guddal

Bernd Ketelsen Striberny

Cathrine Pedersen

Netsanet Gizaw Assefa

Jørn Remi Henriksen

Inge Nilsen

Kersti Överbö

Bjorn Altermark

The invention provides a composition comprising a proteinase or an enzymatically active fragment thereof, said proteinase comprising the amino acid sequence of SEQ ID NO: 1 or comprising an amino acid sequence which is at least about 70% identical to SEQ ID NO: 1, wherein i) the concentration of free calcium in said composition is &#x2264;about 80 &#x3bc;M; or ii) the concentration of monovalent salt in said composition is &#x2265;about 20 mM. Under such conditions, the proteinases and enzymatically active fragments thereof are inducibly thermolabile. The invention further provides samples comprising one or more polypeptides and a proteinase or an enzymatically active fragment thereof, said proteinase comprising the amino acid sequence of SEQ ID NO: 1 or comprising an amino acid sequence which is at least about 70% identical to SEQ ID NO: 1, wherein i) the concentration of free calcium in said sample is &#x2264;about 80 &#x3bc;M; or ii) the concentration of monovalent salt in said sample is &#x2265;about 20 mM. The invention further provides methods comprising the inactivation of such proteinases or enzymatically active fragments thereof, wherein said method comprises the step of heating the sample to inactivate said proteinase or enzymatically active fragment, and wherein i) the concentration of free calcium in said sample is &#x2264;about 80 &#x3bc;M; or ii) the concentration of monovalent salt in said sample is &#x2265;about 20 mM.

Methods of inactivating a proteinase or an enzymatically active fragment thereof

The present invention relates to DNA polymerases. In particular, the present invention relates to DNA polymerases based on a DNA polymerase from asp. The present invention provides an isolated DNA polymerase or an enzymatically active fragment thereof, said DNA polymerase comprising the amino acid sequence of SEQ ID NO:1 or comprising an amino acid sequence which is at least 70% identical to SEQ ID NO:1. The invention also provides nucleic acid molecules comprising a nucleotide sequence that encodes the DNA polymerase. The invention also provides a method of nucleotide polymerisation and a method of amplifying a nucleic acid in which the DNA polymerase or an enzymatically active fragment thereof is used.

Marine DNA polymerases

The invention provides an exonuclease or an enzymatically active fragment thereof, said exonuclease having the amino acid sequence of SEQ ID NO:1 or an amino acid sequence which is at least about 50% identical thereto, wherein said exonuclease or enzymatically active fragment thereof (i) is substantially irreversibly inactivated by heating at a temperature of about 55&#xb0; C. for 10 minutes in a buffer consisting of 10 mM Tris-HCl, pH 8.5 at 25&#xb0; C., 50 mM KCl and 5 mM MgCl; (ii) is substantially specific for single stranded DNA; and (iii) has a 3'-5&#x2032; exonuclease activity. The invention further provides a method of removing single stranded DNA from a sample, a method of nucleic acid amplification, a method of reverse transcription and a method of nucleic acid sequence analysis in which the exonuclease or enzymatically active fragment thereof is used. The invention still further provides nucleic acids encoding said exonuclease or an enzymatically active fragment thereof and kits or compositions comprising the same.

Thermolabile exonucleases

The invention provides an exonuclease or an enzymatically active fragment thereof, said exonuclease having the amino acid sequence of SEQ ID No. 1 or an amino acid sequence which is at least about 50% identical thereto, wherein said exonuclease or enzymatically active fragment thereof (i) is substantially irreversibly inactivated by heating at a temperature of about 55&#xb0; C. for 10 minutes in a buffer consisting of 10 mM Tris-HCI, pH 8.5 at 25&#xb0; C., 50 mM KCI and 5 mM MgCI; (ii) is substantially specific for single stranded DNA; and (iii) has a 3'-5&#x2032; exonuclease activity. The invention further provides a method of removing single stranded DNA from a sample, a method of nucleic acid amplification, a method of reverse transcription and a method of nucleic acid sequence analysis in which the exonuclease or enzymatically active fragment thereof is used. The invention still further provides nucleic acids encoding said exonuclease or an enzymatically active fragment thereof and kits or compositions comprising the same.

The present invention provides an endonuclease I or enzymatically active fragment thereof wherein said endonuclease I has the sequence of SEQ ID No. 4 or a sequence which is at least 70% identical thereto and wherein the amino acid residue which is immediately N-terminal of the FYCGC pentapeptide motif has been substituted with a residue which is negatively charged as well as nucleic acid molecules encoding these enzymes and methods of removing contaminating polynucleotides from a sample using these enzyme.

Endonucleases

The invention provides methods of removing nucleic acid contamination from reverse transcription reactions and hot-start PCR, wherein said hot-start PCR is a barrier hot-start PCR set up and/or involves a hot-start DNA polymerase, which methods comprise use of a DNase that is substantially irreversibly inactivated by heating at a temperature of about 50&#xb0; C. for 5 minutes, and that is substantially specific for double stranded DNA. The invention further provides a DNase that is substantially irreversibly inactivated by heating at a temperature of about 50&#xb0; C. for 5 minutes, and that is substantially specific for double stranded DNA, nucleic acids encoding said DNase and kits or compositions comprising said DNase or said nucleic acid.

Method of removing nucleic acid contamination in reverse transcription and amplification reactions

The present invention relates to a nucleic acid molecule comprising a nucleotide sequence as set forth in SEQ ID Nos. 1 or 20 or having at least 55% sequence identify therewith and/or capable of hybridising under medium stringency conditions to the complementary sequence of SEQ ID Nos. 1 or 20, or the complementary sequence of said sequences, wherein said nucleotide sequence encodes or is complementary to a sequence which encodes a heat labile alkaline phosphatase and to a recombinant heat labile alkaline comprising: (a) all or a significant part of an amino acid sequence as shown in SEQ ID No. 2; or (b) all or a significant part of an amino acid sequence which has at least 60% sequence identify with SEQ ID No. 2 as well as methods for the manufacture thereof. 

Shrimp alkaline phosphatase

It is disclosed a novel enzyme present in cod liver, a DNA sequence encoding the enzyme or operative parts or biologically functional parts thereof, a novel recombinant DNA comprising the gene or the operative or biologically functional parts thereof, a method of preparing the enzyme from cod liver and from bacteria carrying the gene, the bacteria carrying the gene per se, and the use of the protein in monitoring and/or controlling PCR or related reaction systems.

COD URACIL-DNA GLYCOSYLASE, GENE CODING THEREFORE, RECOMBINANT DNA CONTAINING SAID GENE OR OPERATIVE PARTS THEREOF, A METHOD FOR PREPARING SAID PROTEIN AND THE USE OF SAID PROTEIN OR SAID OPERATIVE PARTS THEREOF IN MONITORING OR CONTROLLING PCR

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Tromsø, Norway

Olav Lanes