transformant
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United States of America Patent
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Sep 27, 2016
Grant Date -
Mar 7, 2013
app pub date -
Feb 1, 2011
filing date -
Mar 16, 2010
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Abstract
An object to be solved by the present invention is to provide, for example, a method for producing a Kluyveromyces marxianus transformant by a method for conveniently and efficiently connecting the ends of DNA fragments without using specific restriction enzymes or their recognition sequences, and a method for producing a useful substance using the transformant. The present inventors have found, as means to solve the object, a method comprising introducing two or more linear double-stranded DNA fragments free from a Kluyveromyces marxianus autonomously replicating sequence into Kluyveromyces marxianus and selecting a transformant comprising a desired DNA ligation product with marker gene expression by the desired DNA ligation product as an index, or comprising introducing a linear double-stranded DNA fragment comprising a Kluyveromyces marxianus autonomously replicating sequence, alone or in combination with one or more linear double-stranded DNA fragment(s) different therefrom into Kluyveromyces marxianus and selecting a transformant comprising a desired circular DNA ligation product with marker gene expression by the desired circular DNA ligation product as an index.
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