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    Bacillus subtilis efficiently-induced expression system based on artificial series promoter
    2.
    发明授权
    Bacillus subtilis efficiently-induced expression system based on artificial series promoter 有权

    公开(公告)号：US12098373B2

    公开(公告)日：2024-09-24

    申请号：US17160582

    申请日：2021-01-28

    申请人： Jiangnan University

    发明人： Zhemin Zhou ,  Wenjing Cui ,  Laichuang Han ,  Wenliang Hao ,  Li Zhou ,  Zhongmei Liu ,  Yu Yan

    IPC分类号： C12N15/75

    CPC分类号： C12N15/75 ,  C12N2800/101 ,  C12N2830/002 ,  C12N2830/005 ,  C12N2830/006

    摘要： The present disclosure discloses a Bacillus subtilis efficiently-induced expression system based on an artificial series promoter, and belongs to the technical field of gene engineering. According to the present disclosure, an efficient artificial series constitutive promoter is used, and by compositely designing the promoter and elements (a repressor and a binding site thereof) relevant to an operon, the activity of the constitutive promoter is regulated and controlled by an inducer to finally construct the B. subtilis efficiently-induced expression system induced by the inducer. The result indicates that compared with a P43 strongly constitutive promoter, the activity of the artificial series promoter in the system is about 15 times higher. The activity of the promoter can be accurately controlled by adding different concentrations of inducers. Therefore, the efficient expression system is simple in structure, high in activity and strict in regulating and controlling and has wide application prospects in heterologous protein efficient expression and synthetic biology research.

    Strain producing D-allulose 3-epimerase and application thereof
    5.
    发明授权
    Strain producing D-allulose 3-epimerase and application thereof 有权

    公开(公告)号：US11970723B2

    公开(公告)日：2024-04-30

    申请号：US18187984

    申请日：2023-03-22

    申请人： Jiangnan University

    发明人： Tao Zhang ,  Mengying Hu ,  Bo Jiang ,  Mengli Li

    IPC分类号： C12N9/90 ,  C12N15/75

    CPC分类号： C12N9/90 ,  C12N15/75

    摘要： The present disclosure discloses a strain producing D-allulose 3-epimerase and application thereof, and belongs to the technical field of bioengineering. The present disclosure provides a method for improving the expression of D-allulose 3-epimerase by screening promoters and optimizing RBS thereof. The recombinant Bacillus subtilis constructed using thevectors pP43NMK-hag and pP43NMK-hag-RBS4 provided by the present disclosure improves the enzyme activity of a target gene D-allulose 3-epimerase, and theenzyme activities in shake flasks upon transformation are 1.30 times and 1.69 times that of an original vector. The present disclosure further provides a non-antibiotic resistance vector and a non-antibiotic resistance recombinant B. subtilis strain. Using the non-antibiotic resistance strain B. subtilis 1A751-dal-/pP43NMK-hag-RBS4-dpe-dal provided by the present disclosure, the highest fermentation enzyme activity in a shake flask is 24.72 U/mL, and the enzyme activity in a fermenter is 714.8 U/mL.

    Cas9 variants and methods of use
    7.
    发明授权
    Cas9 variants and methods of use 有权

    公开(公告)号：US11781125B2

    公开(公告)日：2023-10-10

    申请号：US16772910

    申请日：2018-12-11

    申请人： DANISCO USA INC

    发明人： Ryan L. Frisch ,  Hongxian He

    IPC分类号： C12N9/22 ,  C12N15/113 ,  C12N15/70 ,  C12N15/75 ,  C12N15/80

    CPC分类号： C12N9/22 ,  C12N15/113 ,  C12N15/70 ,  C12N15/75 ,  C12N15/80 ,  C12N2310/20 ,  C12N2800/101 ,  C12N2800/80

    摘要： Compositions and methods are provided for variant Cas systems and elements comprising such systems, including, but not limiting to, Cas endonuclease variants, guide polynucleotide/Cas endonuclease complexes comprising Cas endonuclease variants, as well as guide polynucleotides and guide RNA elements that can interact with Cas endonuclease variants. Compositions and methods are provided for genome modification of a target sequence in the genome of a cell. The methods and compositions employ a guide polynucleotide/Cas endonuclease system comprising a Cas9 endonuclease variant to provide an effective system for modifying or altering target sequences within the genome of a cell or organism.