THERMOSTABLE DNA POLYMERASE GENE ISOLATED FROM THERMUS FILIFORMIS AND ITS AMINO ACID SEQUENCE

摘要

PURPOSE: Provided are a thermostable DNA polymerase gene and its amino acid sequence to mass-produce DNA polymerase showing strong stability to heat by cultivating transformants. CONSTITUTION: The thermostable DNA polymerase gene is obtained by the following steps of: i) separating DNA from Thermus filiformis (ATCC 43280¬T); ii) manufacturing a DNA probe for hybridization; iii) implementing agarose gel membrane hybridization; iv) obtaining gene segments of Tfi DNA polymerase from the agarose gel; v) inserting the obtained gene segments of 4.8kb and 2.2kb, into BamH I site of vector pUC18 and transforming E. coli MV1184 with the vector; vi) cultivating the transformants and obtaining two clones, pTFPL and pTFPS; vii) subcloning Tfi DNA segments; and viii) sequencing bases of Tfi DNA.