POLYMERASE ENZYME FROM PYROCOCCUS ABYSSI

1 . A polymerase enzyme according to SEQ ID NO. 1 or any polymerase that shares at least 70%, 80%, 85%, 90%, 95%, 98% amino acid sequence identity thereto, comprising the following mutation(s):

a. at position 409 of SEQ ID NO. 1:

i. serine (S) (L409S) or,

ii. glutamine (Q) (L409Q) or,

iii. tyrosine (Y) (L409Y) or,

iv. phenylalanine (F) (L409F)

b. at position 410 of SEQ ID NO. 1:

i. glycine (G) (Y410G) or,

ii. adenine (A) (Y409A) or,

iii. serine (S) (Y409S),

c. at position 411 of SEQ ID NO. 1:

i. serine (S) (P411S) or,

ii. isoleucine (I) (P411I) or,

iii. cysteine (C) (P411C) or,

iv. adenine (A) (P411A),

wherein the enzyme has little or no 3′-5′ exonuclease activity.

2 . The polymerase enzyme of claim 1 , wherein the polymerase is from an organism belonging to the family of Thermococcaceae, preferably from the genera of Pyrococcus.

3 . The polymerase enzyme according to claim 1 , wherein the polymerase enzyme comprises a L409S mutation, a Y410G mutation and a P411S mutation; and optionally comprises one or more a D141A mutation, a E143A mutation, or a A486L mutation.

4 . The polymerase enzyme according to claim 3 , wherein the polymerase enzyme further comprises the A486L mutation.

5 . The polymerase enzyme according to claim 1 , wherein the polymerase enzyme shares 95% or 98% sequence identity with SEQ ID NO. 1 and comprises the following mutations:

(i) L409S, Y410G, P411S and

(ii) A486L.

6 . The polymerase enzyme according to claim 1 , wherein the polymerase enzyme exhibits an increased rate of incorporation of nucleotides which have been modified at the 3′ sugar hydroxyl such that the substituent is larger in size than the naturally occurring 3′ hydroxyl group, compared to the control polymerase.

7 . A nucleic acid molecule encoding a polymerase enzyme according to claim 1 .

8 . An expression vector comprising the nucleic acid molecule of claim 7 .

9 . A method for incorporating nucleotides which have been modified at the 3′ sugar hydroxyl such that the substituent is larger in size than the naturally occurring 3′ hydroxyl group into DNA comprising the following substances (i) a polymerase enzyme according to claim 1 , (ii) template DNA, (iii) one or more nucleotides, which have been modified at the 3′ sugar hydroxyl such that the substituent is larger in size than the naturally occurring 3′ hydroxyl group.

10 . Use of a polymerase enzyme according to claim 1 for DNA sequencing, DNA labeling, primer extension, amplification or the like.

11 . A kit comprising a polymerase enzyme according to claim 1 .