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THERMOSTABLE DNA POLYMERASE, NUCLEIC ACID SEQUENCE ENCODING IT, VECTOR, HOST CELL, METHOD OF OBTAINING THERMOSTABLE DN POLYMERASE, COMPOSITION FOR DNA SEQUENCING, NUCLEIC ACID SEQUENCING SET AND METHOD OF SEQUENCING NUCLEIC ACID M
THERMOSTABLE DNA POLYMERASE, NUCLEIC ACID SEQUENCE ENCODING IT, VECTOR, HOST CELL, METHOD OF OBTAINING THERMOSTABLE DN POLYMERASE, COMPOSITION FOR DNA SEQUENCING, NUCLEIC ACID SEQUENCING SET AND METHOD OF SEQUENCING NUCLEIC ACID M
The invention provides thermostable DNA polymerase enzymes that comprises the amino acid sequence SerGlnIleXaaLeuArgXaa (SEQ ID NO: 1), wherein "Xaa" at position 4 of this sequence is any amino acid residue but not a glutamic acid residue (Glu), preferably a glycine residue and "Xaa" at position 7 of this sequence is a valine residue (Val) or an isoleucine residue (Ile). The thermostable DNA polymerases of the invention have enhanced efficiency for incorporating unconventional nucleotides, such as ribonucleotides, into DNA products and are advantageous in many in vitro synthesis applications. Such enzymes are particularly useful for use in nucleic acid sequencing protocols and provide novel means for DNA sequence analysis with cost and efficiency advantages. Also claimed are nucleic acids encoding said polymerases, vectors and hoste cells comprising such a nucleic acid, as well as compositions for use in a DNA sequencing reaction, kits and methods for sequencing including such polymerases.
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机译:本发明提供了热稳定的DNA聚合酶,其包含氨基酸序列SerGlnIleXaaLeuArgXaa(SEQ ID NO:1),其中该序列的4位的“ Xaa”是任何氨基酸残基,但不是谷氨酸残基(Glu),优选甘氨酸。残基和该序列第7位的“ Xaa”是缬氨酸残基(Val)或异亮氨酸残基(Ile)。本发明的热稳定DNA聚合酶具有将非常规核苷酸例如核糖核苷酸掺入DNA产物的增强的效率,并且在许多体外合成应用中是有利的。这样的酶特别适用于核酸测序方案,并提供具有成本和效率优势的DNA序列分析的新手段。还要求保护的是编码所述聚合酶的核酸,包含此类核酸的载体和宿主细胞,以及用于DNA测序反应的组合物,包括此类聚合酶的测序试剂盒和方法。
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