To provide a method for preparing a modified protein in which high through-put and efficient function analysis and screening of a modified protein can be established.
(a) A microorganism-origin DNA is prepared from a sample containing the microorganism. (b) Degenerate primers are prepared based on amino acid sequences conserved among first to n-th known proteins having a function of the same type. (c) PCR is performed by using the degenerate primers in the presence of the microorganism-origin DNA. (d) The base sequence of a DNA fragment amplified by the PCR is determined and the amino acid sequence encoded by the DNA fragment is identified. (e) Homology among the amino acid sequence encoded by the DNA fragment and the amino acid sequences of known proteins is evaluated and a known protein having an amino acid sequence showing homology with the amino acid sequence encoded by the DNA fragment is found out. (f) A region encoding the homologous amino acid sequence in the DNA encoding the known protein thus found out in the previous step (e) is substituted with the DNA fragment to thereby prepare a modified DNA. (g) The modified DNA is expressed to prepare a modified protein.
TAKAHASHI MASAAKI
MIYASHITA YUKIKO
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