The patent badge is an abbreviated version of the USPTO patent document. The patent badge does contain a link to the full patent document.

The patent badge is an abbreviated version of the USPTO patent document. The patent badge covers the following: Patent number, Date patent was issued, Date patent was filed, Title of the patent, Applicant, Inventor, Assignee, Attorney firm, Primary examiner, Assistant examiner, CPCs, and Abstract. The patent badge does contain a link to the full patent document (in Adobe Acrobat format, aka pdf). To download or print any patent click here.

Date of Patent:
May. 09, 2023

Filed:

Mar. 24, 2020
Applicant:

National University Corporation Kobe University, Kobe, JP;

Inventors:

Kenji Tsuge, Kobe, JP;

Jun Ishii, Kobe, JP;

Akihiko Kondo, Kobe, JP;

Attorney:
Primary Examiner:
Int. Cl.
CPC ...
C40B 50/06 (2006.01); C12N 15/10 (2006.01); C12N 15/63 (2006.01); C12P 19/34 (2006.01);
U.S. Cl.
CPC ...
C12N 15/1037 (2013.01); C12N 15/63 (2013.01); C12P 19/34 (2013.01); C40B 50/06 (2013.01);
Abstract

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).


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