Yamaguchi University

Babiker Mohamed Ahmed Abdel-Banat

Rinji Akada

Hisashi Hoshida

Jun Asakawa

Sanom Nonklang

An object to be solved by the present invention is to provide, for example, a method for producing atransformant 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 aautonomously replicating sequence intoand 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 aautonomously replicating sequence, alone or in combination with one or more linear double-stranded DNA fragment(s) different therefrom intoand selecting a transformant comprising a desired circular DNA ligation product with marker gene expression by the desired circular DNA ligation product as an index.

[object Object]

It is to provide a noveltransformant having thermotolerance and flocculation property, suitable for the industrial production of bioethanol, by introducing a foreign flocculation gene into, and an efficient method for producing the transformant. The present inventors focused on the flocculation gene FLO ofas a foreign gene to confer flocculation property toand produced a linear DNA fragment comprising a known expression promoter sequence and a FLO gene sequence derived from. As a result of introducing this linear DNA fragment into, the present inventors have confirmed thattransformant can be obtained efficiently, and that the flocculation property of the above transformant is unexpectedly and significantly enhanced. The present invention has been thus completed.

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Ube, Japan

Babiker Mohamed Ahmed Abdel-Banat