| 摘要 |
PROBLEM TO BE SOLVED: To provide a method for efficiently screening thermostable DNA polymerase gene (hereinafter called the gene) which is useful for nucleic-acid amplification reactions for PCR, etc. SOLUTION: This method for codling genes which comprises the following steps, (1) to (8), of: (1) Inserting a mass of gene-containing DNA fragments into a phage vector for in vitro packaging to prepare a library; (2) Infecting host germs with the library, and culturing the germs on a plate to grow plaques; (3) Transferring the plaques on a membrane in surfaces of which a template DNA which becomes a substrate for DNA polymerase has been immobilized; (4) Heat-treating the membrane to which plaques have been transferred. (5) Treating the membrane at temperature of 50-70 deg.C in a reaction liquid in which the labeled dioxynucleoside triphosphate is contained and DNA polymerase functions; (6) Removing excess dioxynicleoside triphoshate which remains in the liquid without being subjected to the reaction; (7) Detecting the labeled dioxynucleoside which has been taken in the membrane surface; and (8) Identifying the clone containing the gene on the basis of the detected signals. |
