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Recombinant His-tagged DNA polymerase. II. Cloning and purification of Thermus aquaticus recombinant DNA polymerase [Stoffel fragment]

100%

Dabrowski S., Kur J.

Acta Biochimica Polonica

1998

tom 45

nr 3

The Stoffel DNA fragment, shortened by 12 bp from 5' end, coding for StoffelDNA polymerase (missing 4 amino acids at N-terminus of Stoffel amino-acids sequence) from the thermophilic Thermus aquaticus (strain YT-1) was amplified, cloned and expressed in Escherichia coli. The recombinant Stoffel fragment contained a polyhistidine tag at the N-terminus (21 additional amino acids) that allowed its single-step isolation by Ni2+ affinity chromatography. The enzyme was characterized and displayed high DNA polymerase activity and thermostability evidently higher than the native Tag DNA polymerase.

Single-stranded DNA-binding proteins [SSBs] - sources and applications in molecular biology

84%

Kur J., Olszewski M., Dlugolecka A., Filipkowski P.

Acta Biochimica Polonica

2005

tom 52

nr 3

Single-stranded DNA-binding proteins (SSBs) play essential roles in DNA replication, recombination, and repair in bacteria, archaea and eukarya. The SSBs share a common core ssDNA-binding domain with a conserved OB (oligonucleotide/oligosaccharide binding) fold. This ssDNA-binding domain was presumably present in the common ancestor to all three major branches of life. In recent years, there has been an increasing interest in SSBs because they are useful for molecular biology methods and for analytical purposes. In this review, we concentrate on recent advances in the discovery of new sources of SSBs as well as certain aspects of their applications in analytical sciences.

Ograniczanie wyników

2 Acta Biochimica Polonica

2 Kur J.

1 Dabrowski S.

1 Dlugolecka A.

1 Filipkowski P.

1 Olszewski M.

1 2005

1 1998

Wyniki wyszukiwania

Recombinant His-tagged DNA polymerase. II. Cloning and purification of Thermus aquaticus recombinant DNA polymerase [Stoffel fragment]

Single-stranded DNA-binding proteins [SSBs] - sources and applications in molecular biology