A thermostable in vitro polymerase complex for template-dependent elongation of nucleic acids in amplification or reverse transcription methods

摘要

A thermostable in vitro complex for template-dependent elongation of nucleic acids comprises a thermostable sliding clamp protein, which is connected with an elongation protein that shows thermostable polymerase activity is new. Independent claims are also included for the following: (1) a thermostable accessory in vitro complex characterized in that it contains a sliding clamp protein and a coupling protein, as defined above; (2) a recombinant DNA sequence, which encodes a thermostable in vitro complex or a thermostable accessory complex; (3) a vector that contains recombinant DNA encoding a sliding clamp protein and a coupling protein and/or an elongation protein; (4) a host cell transformed with one or more vectors of (3); (5) a method for the production of a thermostable in vitro complex or thermostable in vitro accessory complex as above; (6) a method for template-dependent elongation of nucleic acids; (7) a method of marking nucleic acids through generating a single break in the phosphodiester bond of the nucleic acid chain and substituting a nucleotide at the break site with a marked nucleotide with the help of a polymerase, where the polymerase is a thermostable in vitro complex as above; and (8) a reagent kit for elongation and/or amplification and/or reverse-transcription and/or sequencing and/or marking of nucleic acids.