Abstract: The object of the present invention is to provide a method for synthesizing a double-stranded DNA fragment using a PCR method, the method being capable of accurately, easily, efficiently, and quickly synthesizing a double-stranded DNA fragment of interest, regardless of the sequence thereof. The present invention is a method for synthesizing a double-stranded DNA by connecting short double-stranded DNAs to each other by overlap extension PCR to obtain a double-stranded DNA fragment of interest, the method having a plurality of stages of annealing temperature setting in a PCR cycle of the overlap extension PCR.

Abstract: There is provided a method for generating an oil/fat component, in which salt-tolerant algae are cultured in a culture medium of which the salt concentration has been increased in a stepwise manner.

Abstract: The invention provides a method of modifying a targeted site of gram-positive bacterium of a double stranded DNA. The method includes contacting the double-stranded DNA with a complex of a nucleic acid sequence-recognizing module that specifically binds to a target nucleotide sequence in a given double stranded DNA and a nucleic acid base converting enzyme to convert, delete, or insert one or more nucleotides in the targeted site without cleaving at least one strand of the double stranded DNA in the targeted site, by introducing the nucleic acid encoding the complex into the gram-positive bacterium. The invention also provide a nucleic acid-modifying enzyme complex of a nucleic acid sequence-recognizing module that specifically binds to a target nucleotide sequence in a double stranded DNA of a gram-positive bacterium and a nucleic acid base converting enzyme bonded to each other, which complex is used for the method.

Abstract: The present invention provides a method of modifying a targeted site of a double stranded DNA of a monocot cell, comprising a step of contacting a complex wherein a nucleic acid sequence-recognizing module that specifically binds to a target nucleotide sequence in the given double stranded DNA and a nucleic acid base converting enzyme are bonded, with said double stranded DNA, to convert one or more nucleotides in the targeted site to other one or more nucleotides or delete one or more nucleotides, or insert one or more nucleotides into said targeted site, without cleaving at least one strand of said double stranded DNA in the targeted site, wherein the double stranded DNA is contacted with the complex by introducing a nucleic acid encoding the complex into the monocot cell.

Abstract: There is provided a method for generating an oil/fat component by means of culturing algae, in which marine algae belonging to chlamydomonas are cultured in a culture medium containing sea salt.

Abstract: The present invention provides a method of modifying a targeted site of a double stranded DNA in a host cell, the method including introducing (a) a DNA encoding a crRNA containing a sequence complementary to a target strand of a target nucleotide sequence in the given double stranded DNA, and (b) a DNA encoding a protein group constituting Cascade and a nucleic acid base converting enzyme, in which the nucleic acid base converting enzyme is constituted in a form capable of forming a complex with any protein in the protein group, into the host cell to convert one or more nucleotides in the targeted site to other one or more nucleotides, or delete one or more nucleotides, or insert one or more nucleotides into said targeted site, without cleaving the double stranded DNA in the targeted site.

Abstract: The production method of the present invention includes: a first fermentation step of adding at least one microorganism selected from yeast, Escherichia coli, and bacteria of the genus Corynebacterium to a first concentrated sugar solution to ferment the first concentrated sugar solution and subjecting the resulting first fermented solution to solid-liquid separation, the first concentrated sugar solution being obtained by using a nanofiltration membrane and/or a reverse osmosis membrane to concentrate a sugar solution obtained using a herbaceous plant of the family Gramineae or Cucurbitaceae as a raw material; and a second fermentation step of adding a second concentrated sugar solution to a solid component obtained in the first fermentation step to ferment the second concentrated sugar solution with the microorganism used in the first fermentation step, and subjecting the resulting second fermented solution to solid-liquid separation.

Abstract: The present invention aims to provide a yeast that is genetically modified so as to more highly produce glutathione, and a method of producing glutathione utilizing the yeast. The present invention provides a method of producing glutathione, including culturing a yeast whose thiol oxidase activity is increased as compared to the parent strain in a culture medium to produce glutathione, and recovering glutathione from the cultured broth obtained.

Abstract: The present invention provides a method of modifying a targeted site of a double stranded DNA, including a step of contacting a complex wherein a nucleic acid sequence-recognizing module that specifically binds to a target nucleotide sequence in a selected double stranded DNA and DNA glycosylase with sufficiently low reactivity with a DNA having an unrelaxed double helix structure (unrelaxed DNA) are bonded, with the double stranded DNA, to convert one or more nucleotides in the targeted site to other one or more nucleotides or delete one or more nucleotides, or insert one or more nucleotides into the targeted site, without cleaving at least one strand of the double stranded DNA in the targeted site.

Abstract: Disclosed is a polynucleotide for cell surface expression. The polynucleotide of the invention comprises a promoter, a secretion signal sequence, a sequence encoding an intended protein, and a sequence encoding a cell surface-localized protein or a cell membrane-binding domain thereof, wherein the promoter is a promoter of a gene encoding the cell surface-localized protein. Provided is a polynucleotide for cell surface expression allowing the production of yeast displaying an enzyme on the cell surface with a high activity.

Abstract: An expression vector is disclosed which contains a promoter DNA; a DNA encoding a peptide having a defined amino acid sequence and having secretion signal activity; and a DNA encoding an intended protein or a cloning site for insertion of the DNA encoding an intended protein. An expression vector is also disclosed which contains a promoter DNA; a DNA encoding any peptide having a defined amino acid sequence and having secretion signal activity; a DNA encoding an intended protein or a cloning site for insertion of the DNA encoding an intended protein; and a DNA encoding an anchor domain. The peptide having secretion signal activity allows for secretory production and cell surface display of a protein with high activity, in yeast.

Abstract: The present invention provides a method of modifying a targeted site of a double stranded DNA, including a step of contacting a complex wherein a nucleic acid sequence-recognizing module that specifically binds to a target nucleotide sequence in a selected double stranded DNA and a nucleic acid base converting enzyme are bonded, with the double stranded DNA, to convert one or more nucleotides in the targeted site to other one or more nucleotides or delete one or more nucleotides, or insert one or more nucleotides into the targeted site, without cleaving at least one chain of the double stranded DNA in the targeted site.

Abstract: The present invention provides a method for producing a useful substance efficiently from cellulose without using any cellulase preparation. According to the disclosures of the present specification, in the production of a useful substance from a cellulose-containing material, the cellulose-containing material is brought into contact with an ionic liquid to cause the ionic liquid to permeate the cellulose-containing material, and a carbon source comprising the cellulose in the cellulose-containing material is simultaneously saccharified and fermented by a cellulase-producing microorganism in the presence of the ionic liquid.

Abstract: A method for producing ethanol from lignocellulosic biomass using yeast at low cost is provided. The method of the present invention for producing ethanol from lignocellulosic biomass includes steps of (1) pretreating lignocellulosic biomass, (2) treating a cellulose fraction obtained in Step (1) with a cellulose hydrolase, (3) mixing saccharified biomass obtained in Step (2) with yeasts to perform ethanol fermentation, and (4) subjecting a fermentation product obtained in Step (3) to a solid-liquid separation, wherein a cycle consisting of Steps (1), (2), (3) and (4) is repeated twice or more, and yeasts obtained in Step (4) are used as all or a portion of yeasts in Step (3) of the subsequent cycle.

Abstract: Disclosed is a polynucleotide for cell surface expression. The polynucleotide of the invention comprises a promoter, a secretion signal sequence, a sequence encoding an intended protein, and a sequence encoding a cell surface-localized protein or a cell membrane-binding domain thereof, wherein the promoter is a promoter of a gene encoding the cell surface-localized protein. Provided is a polynucleotide for cell surface expression allowing the production of yeast displaying an enzyme on the cell surface with a high activity.

Abstract: Provided is a method for efficiently producing ethanol by ethanol fermentation from xylose using a saccharified biomass, which contains various fermentation inhibitors. A method for producing ethanol from biomass of the present invention includes the step of culturing a xylose-utilizing yeast transformed so as to overexpress a gene for an acetic acid-responsive transcription factor in combination with a saccharified biomass.

Abstract: There is provided a method for generating an oil/fat component, in which salt-tolerant algae are cultured in a culture medium of which the salt concentration has been increased in a stepwise manner.

Abstract: There is provided a method for generating an oil/fat component by means of culturing algae, in which marine algae belonging to Chlamydomonas are cultured in a culture medium containing sea salt.

Abstract: The present invention provides a method for measuring oocyst of protozoa, such as Cryptosporidium, in an environment sample with high sensitivity at low cost within a short period of time; and a detecting reagent for use therein. Magnetic fine particles of 5 to 500 nm particle diameter having, immobilized thereto, binding factors for specific recognition of oocyst are added to an analyte containing a protozoan oocyst to form oocyst/binding factor/magnetic fine particle complexes by using a binding reaction to the oocyst, the formed complexes are recovered by a magnetic separation, and the protozoan oocysts contained in the complexes are assayed. Further, there is provided, for conducting the above method, a reagent for detecting protozoan oocysts comprising magnetic fine particles of 5 to 500 nm particle diameter having, immobilized thereto, antibodies against oocysts or binding factors for recognizing the antibodies.

Abstract: An object of the present invention is to provide a method for producing ethanol efficiently even in the presence of a fermentation inhibitor in a saccharified biomass. The present invention provides a method for producing ethanol from biomass, comprising: culturing a transformed xylose-utilizing yeast to overexpress the gene for at least one pentose phosphate pathway metabolic enzyme, with a saccharified biomass.