Yamaguchi University

Toyota Jidosha Kabushiki Kaisha

Jx Nippon Oil Energy Corporation

Kohjin Life Sciences Co., Ltd.

Mitsubishi Corporation Life Sciences Limited

Rinji Akada

Hisashi Hoshida

Satoshi Katahira

Kenro Tokuhiro

Mikiko Nakamura

Noriko Shisa

Kozue Mutaguchi

Takeshi Uemura

Babiker Mohamed Ahmed Abdel-Banat

Jun Asakawa

Masamitsu Ide

Sanom Nonklang

In order to provide a culture medium eliminating variation by lot of component concentrations, the present invention provides a culture medium, and preferably a chemical-synthetic culture medium, imparting a proliferation ability to yeast that is equivalent to or greater than that of a YPD culture medium. A culture medium is provided which includes sugars as a carbon source capable of being assimilated by yeast; amino acids as a nitrogen source; vitamins; inositol; zinc ion (Zn); potassium ion (K), and magnesium ion (Mg), and in which inositol concentration is between 50 and 10,000 mg/L.

Medium for yeasts

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.

Provided is an efficiency improving agent for gene transfer to mammalian cells, a method for improving efficiency of gene transfer to mammalian cells, and a method for transforming mammalian cells. The method is characterized in that tRNA is used in combination with a lipofection reagent. Preferably, the agent may be used so that the tRNA concentration in a lipofection solution falls within the range of 3 to 50 &#x3bc;g/mL, and the concentration in a culture is approximately 1/10. More preferably, tRNA and PEG may be used in combination with a lipofection reagent. According to the present invention, gene transfer to mammalian cells with high efficiency can be achieved.

Agent for improving gene transfer efficiency to mammalian cells

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

Yamaguchi, Japan

Rinji Akada