Yamaguchi University

Toyota Jidosha Kabushiki Kaisha

Jx Nippon Oil Energy Corporation

Hisashi Hoshida

Rinji Akada

Satoshi Katahira

Kenro Tokuhiro

Noriko Shisa

Kozue Mutaguchi

Takeshi Uemura

Babiker Mohamed Ahmed Abdel-Banat

Mikiko Nakamura

Jun Asakawa

Masamitsu Ide

Sanom Nonklang

Plasmid vectors have been widely used as a carrier of a DNA sequence capable of expressing a target RNA in cells. However, construction of these plasmid vectors requires technical skill and time. Thus, a quicker and easier method is required therefor. To solve this problem, a method using a linear DNA that has been amplified by the PCR method is examined. However, this method is disadvantageous in that RNA expression in cells is extremely low. Under these circumstances, the present inventors attempted to develop an RNA expression method using a linear DNA which can be produced mainly by using the PCR method alone and which enables a high level of RNA expression. As the results of intensive studies on terminator sequences to be used in a linear DNA, the present inventors found a smallest unit of a terminator sequence enabling linear DNA expression equivalent to that when using a plasmid vector. A linear DNA including the aforesaid terminator sequence can be produced quickly and easily, and enables RNA expression at a higher level. The present invention has been completed based on these findings.

Terminator sequence-containing reverse primer for overexpression and linear DNA

The promoter of the present invention causes a desired gene to be highly expressed, especially in thermotolerant yeast. The promoter is located upstream of the PIR1 gene or the CTR1 gene on thechromosome and comprises a region controlling expression of the PIR1 gene or the CTR1 gene.

Promoter and use thereof

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]

A yeast of the genusis modified to improve the ethanol yield from xylose by attenuation of at least one gene selected from the group consisting of the ADH1 gene derived from, a gene functionally equivalent to the ADH1 gene, the ADH4 gene derived from, and a gene functionally equivalent to the ADH4 gene.

Yeast mutant of kluyveromyces and method for ethanol production using the same

Provided is a novel high-expression promoter, namely a GAL1 promoter, derived from. Also provided are the following, characterized by the use of the provided high-expression promoter; a recombinant polynucleotide containing said high-expression promoter; a vector containing said recombinant polynucleotide; a transformant obtained by introducing said recombinant polynucleotide or vector into yeast; a method using said transformant for high expression of a target gene; and a method using said transformant to manufacture the gene product of a target gene.

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.

Growing community of inventors

Ube, Japan

Hisashi Hoshida