序号 专利名 申请号 申请日 公开(公告)号 公开(公告)日 发明人
1 合成遺伝子の設計方法 JP2014185629 2014-09-11 JP2016054715A 2016-04-21 井上 敏
【課題】“使用頻度の高いコドンのみを選択的に使用し、かつGC含量が60%以上”である遺伝子設計法が高発現タンパク質の一般的ルールとなり得るか否かの検証方法の提供。
【解決手段】目的タンパク質遺伝子の塩基配列を、ヒト細胞で使用頻度の高いコドンのみを選択し、かつGC含量が60%以上になるよう改変する工程を含む、選択型遺伝子の設計方法、及び、発光タンパク質または発光酵素が、イクオリン、クライチィンII、ガウシアルシフェラーゼ、エビルシフェラーゼの発光触媒タンパク質の変異体、北米産ホタルルシフェラーゼ、ゲンジボタルルシフェラーゼ、またはレニラルシフェラーゼから選択される選択型遺伝子、及び、選択型遺伝子を用いたタンパク質の製造方法
【選択図】なし
2 Codon-optimized gene for mutated shrimp luciferase and method for use thereof US15291367 2016-10-12 US10006009B2 2018-06-26 Satoshi Inouye; Junichi Sato
There has been a demand for a codon-optimized gene for the mutated catalytic domain of Oplophorus luciferase, which is capable of efficiently expressing a protein both in a cultured animal cell and Escherichia coli. There has also been a demand for a substrate coelenterazine analog showing a higher activity than that of native 19 kDa protein. The invention provides a polynucleotide comprising a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 2. According to the invention, bis-coelenterazine is used as a substrate coelenterazine analog suitable for the photoprotein encoded by the polynucleotide comprising the polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 2.
3 Mutated genes for the catalytic protein of oplophorus luciferase and use thereof US14685643 2015-04-14 US09868941B2 2018-01-16 Satoshi Inouye; Yuiko Miura; Junichi Sato
A novel luciferase that is distinct from conventional luciferase has been desired. A luciferase mutant comprising an amino acid sequence in which glutamic acid at position 4, arginine at position 11, leucine at position 18 and valine at position 27 are substituted with other amino acids, in the amino acid sequence of SEQ ID NO: 2.
4 Process for production of recombinant proteins as a soluble form US14476624 2014-09-03 US09738692B2 2017-08-22 Satoshi Inouye; Yuiko Sahara
A target protein is prepared as soluble protein using a recombinant protein expression system. An expression vector is used that includes (1) an expression-inducible promoter sequence; (2) a first coding sequence including a polynucleotide coding for a polypeptide that is represented by the formula (Z)n; and (3) a second coding sequence that includes a polynucleotide that codes for a target protein. A method of producing the target protein is also used that includes expressing protein using this expression vector.
5 CODON-OPTIMIZED GENE FOR MUTATED SHRIMP LUCIFERASE AND METHOD FOR USE THEREOF US15191730 2016-06-24 US20160298171A1 2016-10-13 Satoshi INOUYE; Junichi SATO
There has been a demand for a codon-optimized gene for the mutated catalytic domain of Oplophorus luciferase, which is capable of efficiently expressing a protein both in a cultured animal cell and Escherichia coli. There has also been a demand for a substrate coelenterazine analogue showing a higher activity than that of native 19 kDa protein. The invention provides a polynucleotide comprising a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 2. According to the invention, bis-coelenterazine is used as a substrate coelenterazine analogue suitable for the photoprotein encoded by the polynucleotide comprising the polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 2.
6 MUTATED GENES FOR THE CATALYTIC PROTEIN OF OPLOPHORUS LUCIFERASE AND USE THEREOF US14685643 2015-04-14 US20150299675A1 2015-10-22 Satoshi INOUYE; Yuiko MIURA; Junichi SATO
A novel luciferase that is distinct from conventional luciferase has been desired. A luciferase mutant comprising an amino acid sequence in which glutamic acid at position 4, arginine at position 11, leucine at position 18 and valine at position 27 are substituted with other amino acids, in the amino acid sequence of SEQ ID NO: 2.
7 COELENTERAZINE ANALOGUES US15982514 2018-05-17 US20180334463A1 2018-11-22 Mary Hall; Thomas Kirkland; Thomas Machleidt; Anton Shakhmin; Joel R. Walker
Described are coelenterazine analogues, methods for making the analogues, kits comprising the analogues, and methods of using the compounds for the detection of luminescence in luciferase-based assays.
8 DESIGN METHOD FOR SYNTHETIC GENES US15905521 2018-02-26 US20180171356A1 2018-06-21 Satoshi Inouye
The present invention provides a method of designing an optimized gene which comprises altering a nucleotide sequence of a target protein gene, so that only preferential codons with high frequency of use in human cells are selected and a GC content of not less than 60% is achieved. A gene design method which involves the feature “only preferential codons with high frequency of use are selected and a GC content of not less than 60% is achieved” can be established as a general rule for preparing proteins with high expression level, in order to obtain chemically synthesized genes for proteins capable of high-level expression in eukaryotes.
9 Coelenterazine analogues US15661582 2017-07-27 US10000500B2 2018-06-19 Mary Hall; Thomas Kirkland; Thomas Machleidt; Anton Shakhmin; Joel R. Walker
Described are coelenterazine analogs, methods for making the analogs, kits comprising the analogs, and methods of using the compounds for the detection of luminescence in luciferase-based assays.
10 Method for synthetic genes US14848535 2015-09-09 US09938539B2 2018-04-10 Satoshi Inouye
The present invention provides a method of designing an optimized gene which comprises altering a nucleotide sequence of a target protein gene, so that only preferential codons with high frequency of use in human cells are selected and a GC content of not less than 60% is achieved. A gene design method which involves the feature “only preferential codons with high frequency of use are selected and a GC content of not less than 60% is achieved” can be established as a general rule for preparing proteins with high expression level, in order to obtain chemically synthesized genes for proteins capable of high-level expression in eukaryotes.
11 COELENTERAZINE ANALOGUES US15661582 2017-07-27 US20180030059A1 2018-02-01 Mary Hall; Thomas Kirkland; Thomas Machleidt; Anton Shakhmin; Joel R. Walker
Described are coelenterazine analogues, methods for making the analogues, kits comprising the analogues, and methods of using the compounds for the detection of luminescence in luciferase-based assays.
12 Codon-optimized gene for mutated shrimp luciferase and method for use thereof US15191730 2016-06-24 US09506104B2 2016-11-29 Satoshi Inouye; Junichi Sato
There has been a demand for a codon-optimized gene for the mutated catalytic domain of Oplophorus luciferase, which is capable of efficiently expressing a protein both in a cultured animal cell and Escherichia coli. There has also been a demand for a substrate coelenterazine analog showing a higher activity than that of native 19 kDa protein. The invention provides a polynucleotide comprising a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 2. According to the invention, bis-coelenterazine is used as a substrate coelenterazine analog suitable for the photoprotein encoded by the polynucleotide comprising the polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 2.
13 CODON-OPTIMIZED GENE FOR MUTATED SHRIMP LUCIFERASE AND METHOD FOR USE THEREOF US14188838 2014-02-25 US20140242574A1 2014-08-28 Satoshi INOUYE; Junichi SATO
There has been a demand for a codon-optimized gene for the mutated catalytic domain of Oplophorus luciferase, which is capable of efficiently expressing a protein both in a cultured animal cell and Escherichia coli. There has also been a demand for a substrate coelenterazine analogue showing a higher activity than that of native 19 kDa protein. The invention provides a polynucleotide comprising a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 2. According to the invention, bis-coelenterazine is used as a substrate coelenterazine analogue suitable for the photoprotein encoded by the polynucleotide comprising the polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 2.
14 エビルシフェラーゼの触媒蛋白質のキメラ遺伝子及びその使用法 JP2014047379 2014-03-11 JP6256118B2 2018-01-10 井上 敏; 佐藤 淳一
15 エビルシフェラーゼの触媒蛋白質の変異遺伝子とその使用法 JP2014084499 2014-04-16 JP2015202096A 2015-11-16 井上 敏; 三浦 由依子; 佐藤 淳一
【課題】深海エビであるオプロフォーラス(Oplophorus gracilirostris)由来のルシフェラーゼの細胞外への分泌型の新規変異体の提供。
【解決手段】オプロフォーラスルシフェラーゼにおいて発光を触媒する分子量19kDa蛋白質の既知の変異体(nanoKAZ)の変異部位について検証をおこない、変異の選択及び組合せによりセレンテラジンを発光基質としてより活性が高く、動物培養細胞内で発現したときに真核細胞生物由来の分泌シグナル配列の有無にかかわらず細胞外へ分泌するルシフェラーゼ変異体。
【選択図】なし
16 MUTATED GENES FOR THE CATALYTIC PROTEIN OF OPLOPHORUS LUCIFERASE AND USE THEREOF US15834147 2017-12-07 US20180119112A1 2018-05-03 Satoshi INOUYE; Yuiko MIURA; Junichi SATO
A novel luciferase that is distinct from conventional luciferase has been desired. A luciferase mutant comprising an amino acid sequence in which glutamic acid at position 4, arginine at position 11, leucine at position 18 and valine at position 27 are substituted with other amino acids, in the amino acid sequence of SEQ ID NO: 2.
17 CODON-OPTIMIZED GENE FOR MUTATED SHRIMP LUCIFERASE AND METHOD FOR USE THEREOF US15291367 2016-10-12 US20170029789A1 2017-02-02 Satoshi INOUYE; Junichi SATO
There has been a demand for a codon-optimized gene for the mutated catalytic domain of Oplophorus luciferase, which is capable of efficiently expressing a protein both in a cultured animal cell and Escherichia coli. There has also been a demand for a substrate coelenterazine analogue showing a higher activity than that of native 19 kDa protein. The invention provides a polynucleotide comprising a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 2. According to the invention, bis-coelenterazine is used as a substrate coelenterazine analogue suitable for the photoprotein encoded by the polynucleotide comprising the polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 2.
18 Codon-optimized gene for mutated shrimp luciferase and method for use thereof US15067591 2016-03-11 US09404145B2 2016-08-02 Satoshi Inouye; Junichi Sato
There has been a demand for a codon-optimized gene for the mutated catalytic domain of Oplophorus luciferase, which is capable of efficiently expressing a protein both in a cultured animal cell and Escherichia coli. There has also been a demand for a substrate coelenterazine analog showing a higher activity than that of native 19 kDa protein. The invention provides a polynucleotide comprising a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 2. According to the invention, bis-coelenterazine is used as a substrate coelenterazine analog suitable for the photoprotein encoded by the polynucleotide comprising the polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 2.
19 CODON-OPTIMIZED GENE FOR MUTATED SHRIMP LUCIFERASE AND METHOD FOR USE THEREOF US15067591 2016-03-11 US20160186234A1 2016-06-30 Satoshi INOUYE; Junichi SATO
There has been a demand for a codon-optimized gene for the mutated catalytic domain of Oplophorus luciferase, which is capable of efficiently expressing a protein both in a cultured animal cell and Escherichia coli. There has also been a demand for a substrate coelenterazine analogue showing a higher activity than that of native 19 kDa protein. The invention provides a polynucleotide comprising a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 2. According to the invention, bis-coelenterazine is used as a substrate coelenterazine analogue suitable for the photoprotein encoded by the polynucleotide comprising the polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 2.
20 Codon-optimized gene for mutated shrimp luciferase and method for use thereof US14188838 2014-02-25 US09315783B2 2016-04-19 Satoshi Inouye; Junichi Sato
There has been a demand for a codon-optimized gene for the mutated catalytic domain of Oplophorus luciferase, which is capable of efficiently expressing a protein both in a cultured animal cell and Escherichia coli. There has also been a demand for a substrate coelenterazine analogue showing a higher activity than that of native 19 kDa protein. The invention provides a polynucleotide comprising a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 2. According to the invention, bis-coelenterazine is used as a substrate coelenterazine analogue suitable for the photoprotein encoded by the polynucleotide comprising the polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 2.
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