The purified enzyme termoestavel, encoder interface gene of enzyme, plasmidio selected, bactereofogo, structure of the enzyme is stable.Process for amplifying at least a sequence specific nucleic acid, a process to detect the presence or absence of at least one sequence specific nucleic acid.Process for detecting the presence or absence of at least one change of the nucleotideos in sequence in one or more nucleic acids.Process for cloning vector for cloning one or more specific sequences of nucleic acid and nucleic acid sequence amplified

摘要

A purified thermostable enzyme is obtained that has unique characteristics. Preferably the enzyme is isolated from the Thermus aquaticus species and has a molecular weight of about 86,000-90,000 daltons. The thermostable enzyme may be native or recombinant and may be used in a temperature-cycling chain reaction wherein at least one nucleic acid sequence is amplified in quantity from an existing sequence with the aid of selected primers and nucleotide triphosphates. The amplification process comprises treating separate complementary strands of the nucleic acid with a molar excess of two oligonucleotide primers, extending the primers with a thermostable enzyme to form complementary primer extension products which act as templates for synthesizing the desired nucleic acid sequence, and detecting the sequence so amplified. The steps of the reaction can be repeated as often as desired and involve temperature cycling to effect hybridization, promotion of activity of the enzyme, and denaturation of the hybrids formed. The enzyme is preferably stored in a buffer of non-ionic detergents that lends stability to the enzyme.