Kobe University

Kaneka Corporation

Jun Ishii

Akihiko Kondo

Yasuyuki Nakamura

Teruyuki Nishi

Kenji Tsuge

Yoichiro Ito

The present invention addresses the problem of providing a novel method which is for preparing a DNA fragment for microbial cell transformation, and by which the combinatorial library of a long-chain DNA can be efficiently constructed and confirmation of the genotype of the obtained clone is facilitated. The present invention is a method for preparing a DNA fragment, which is for microbial cell transformation and has at least one insert DNA unit that includes a DNA containing an effective replication origin in a host microorganism and an insert DNA in which unit DNAs are linked, the method being characterized by including: (A) a step for preparing, through an OGAB method, a plurality of types of plasmids having an insert DNA unit in which a plurality of types of unit DNAs capable of being linked in a specific linking order are linked; (B) a step for decomposing a plasmid into unit DNAs by treating the plurality of types of plasmids prepared in the step (A) with a restriction enzyme suitable for each plasmid and preparing a mixed liquid of a plurality of types of unit DNAs; and (C) a step for preparing a long-chain DNA fragment by re-assembling the unit DNAs through the OGAB method by using the mixed liquid of a plurality of types of unit DNAs obtained in the step (B).

Method for constructing chimeric plasmid library

One or more embodiments of the present invention are to provide a new means of improving the productivity of a target protein. The present inventors have identified a novel protein consisting of the amino acid sequence set forth in SEQ ID NO: 2 through an exhaustive analysis of the nucleotide sequence of chromosomal DNA of a yeast belonging to the genus. Activation of a gene encoding the novel protein provides a cell having an improved productivity of a target protein.

Cell and method for producing target protein using same

Growing community of inventors

Hyogo, Japan

Jun Ishii