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    METHOD FOR PRODUCING HUMAN EPIDERMAL GROWTH FACTOR IN LARGE VOLUME FROM YEAST
    3.
    发明申请
    METHOD FOR PRODUCING HUMAN EPIDERMAL GROWTH FACTOR IN LARGE VOLUME FROM YEAST 有权
    从日本大批量生产人类生殖生长因子的方法

    公开(公告)号：US20130323785A1

    公开(公告)日：2013-12-05

    申请号：US13883264

    申请日：2011-11-04

    Applicant: Jung Hoon Sohn ,  Jung Hoon Bae ,  Mi Jin Kim ,  Hyun Jin Kim ,  Soon Ho Park ,  Kwang Mook Lim

    Inventor： Jung Hoon Sohn ,  Jung Hoon Bae ,  Mi Jin Kim ,  Hyun Jin Kim ,  Soon Ho Park ,  Kwang Mook Lim

    IPC: C07K14/485 ,  C12R1/645

    CPC classification number: C07K14/485 ,  C12N15/81 ,  C12R1/645

    Abstract: The present invention relates to a method for producing hEGF (human epidermal growth factor) which has the same activity as the wild form, in high concentration and with a high degree of purity. More specifically, the invention relates to an hEGF expression vector comprising a nucleic acid sequence coding for the polypeptide of sequence number 14; a host cell in which the expression vector has been genetically transformed; and a method for producing hEGF, comprising a step in which the expression vector is created and is genetically transformed in yeast from which the KEX1 gene is lacking. Using the method of the present invention, it is possible to produce a large volume of human derived EGF which has the same size and activity as human derived EGF, and this EGF can be used in various ways such as in medicine and cosmetics.

    Abstract translation: 本发明涉及以高浓度和高纯度生产具有与野生型相同的活性的hEGF（人表皮生长因子）的方法。 更具体地，本发明涉及包含编码序列号14的多肽的核酸序列的hEGF表达载体; 其中表达载体已被遗传转化的宿主细胞; 以及生产hEGF的方法，包括其中缺少表达载体并在KEX1基因缺失的酵母中遗传转化的步骤。 使用本发明的方法，可以生产与人源性EGF具有相同大小和活性的大量人源性EGF，并且该EGF可以以各种方式用于医药和化妆品中。

    Genome walking method for cloning of unknown DNA sequences adjacent to known sequences
    7.
    发明授权
    Genome walking method for cloning of unknown DNA sequences adjacent to known sequences 有权
    用于克隆与已知序列相邻的未知DNA序列的基因组步行方法

    公开(公告)号：US09260749B2

    公开(公告)日：2016-02-16

    申请号：US13345616

    申请日：2012-01-06

    Applicant: Jung Hoon Sohn ,  Jung Hoon Bae

    Inventor： Jung Hoon Sohn ,  Jung Hoon Bae

    IPC: C12Q1/68

    CPC classification number: C12Q1/6855 ,  C12Q2525/186

    Abstract: A method and a kit for cloning a nucleotide sequence adjacent to a known nucleotide sequence by PCR are disclosed. The method includes 1) preparing a DNA fragment with cohesive ends by cleaving a DNA that contains the known first nucleotide sequence and the second nucleotide sequence adjacent thereto using a restriction enzyme; 2) modifying the 3′ end of the DNA fragment with a nucleotide analog to block further elongation of the DNA fragment; 3) linking the cohesive ends of the 3′ end-modified DNA fragment with a linker, adapter or cassette having a cohesive end sequence complementary thereto; and 4) performing PCR using a known first nucleotide sequence-specific primer, and a linker, adapter or cassette sequence-specific primer.

    Abstract translation: 公开了通过PCR克隆与已知核苷酸序列相邻的核苷酸序列的方法和试剂盒。 该方法包括：1）通过使用限制酶切割包含已知第一个核苷酸序列的DNA和与其相邻的第二个核苷酸序列，制备具有粘性末端的DNA片段; 2）用核苷酸类似物修饰DNA片段的3'末端以阻断DNA片段的进一步延伸; 3）将3'末端修饰的DNA片段的粘性末端与具有与其互补的内聚端序列的接头，接头或盒连接; 和4）使用已知的第一核苷酸序列特异性引物和接头，衔接子或盒序列特异性引物进行PCR。

    Method for producing human epidermal growth factor in large volume from yeast
    8.
    发明授权
    Method for producing human epidermal growth factor in large volume from yeast 有权
    从酵母中大量生产人表皮生长因子的方法

    公开(公告)号：US08986956B2

    公开(公告)日：2015-03-24

    申请号：US13883264

    申请日：2011-11-04

    Applicant: Jung Hoon Sohn ,  Jung Hoon Bae ,  Mi Jin Kim ,  Hyun Jin Kim ,  Soon Ho Park ,  Kwang Mook Lim

    Inventor： Jung Hoon Sohn ,  Jung Hoon Bae ,  Mi Jin Kim ,  Hyun Jin Kim ,  Soon Ho Park ,  Kwang Mook Lim

    IPC: C12P21/04 ,  C12N1/16 ,  C12N15/09 ,  C07H21/04 ,  C07K14/485 ,  C12N15/81 ,  C12R1/645

    CPC classification number: C07K14/485 ,  C12N15/81 ,  C12R1/645

    Abstract: The present invention relates to a method for producing hEGF (human epidermal growth factor) which has the same activity as the wild form, in high concentration and with a high degree of purity. More specifically, the invention relates to an hEGF expression vector comprising a nucleic acid sequence coding for the polypeptide of sequence number 14; a host cell in which the expression vector has been genetically transformed; and a method for producing hEGF, comprising a step in which the expression vector is created and is genetically transformed in yeast from which the KEX1 gene is lacking. Using the method of the present invention, it is possible to produce a large volume of human derived EGF which has the same size and activity as human derived EGF, and this EGF can be used in various ways such as in medicine and cosmetics.

    Abstract translation: 本发明涉及以高浓度和高纯度生产具有与野生型相同的活性的hEGF（人表皮生长因子）的方法。 更具体地，本发明涉及包含编码序列号14的多肽的核酸序列的hEGF表达载体; 其中表达载体已被遗传转化的宿主细胞; 以及生产hEGF的方法，包括其中缺少表达载体并在KEX1基因缺失的酵母中遗传转化的步骤。 使用本发明的方法，可以生产与人源性EGF具有相同大小和活性的大量人源性EGF，并且该EGF可以以各种方式用于医药和化妆品中。

    NOVEL GENOME WALKING METHOD FOR CLONING OF UNKNOWN DNA SEQUENCES ADJACENT TO KNOWN SEQUENCE
    10.
    发明申请
    NOVEL GENOME WALKING METHOD FOR CLONING OF UNKNOWN DNA SEQUENCES ADJACENT TO KNOWN SEQUENCE 有权
    用于克隆与已知序列相关的未知DNA序列的新基因组方法

    公开(公告)号：US20120171727A1

    公开(公告)日：2012-07-05

    申请号：US13345616

    申请日：2012-01-06

    Applicant: Jung Hoon SOHN ,  Jung Hoon BAE

    Inventor： Jung Hoon SOHN ,  Jung Hoon BAE

    IPC: C12P19/34 ,  C09K3/00

    CPC classification number: C12Q1/6855 ,  C12Q2525/186

    Abstract: A method and a kit for cloning a nucleotide sequence adjacent to a known nucleotide sequence by PCR are disclosed. The method includes 1) preparing a DNA fragment with cohesive ends by cleaving a DNA that contains the known first nucleotide sequence and the second nucleotide sequence adjacent thereto using a restriction enzyme; 2) modifying the 3′ end of the DNA fragment with a nucleotide analogue to block further elongation of the DNA fragment; 3) linking the cohesive ends of the 3′ end-modified DNA fragment with a linker, adapter or cassette having a cohesive end sequence complementary thereto; and 4) performing PCR using a known first nucleotide sequence-specific primer, and a linker, adapter or cassette sequence-specific primer.

    Abstract translation: 公开了通过PCR克隆与已知核苷酸序列相邻的核苷酸序列的方法和试剂盒。 该方法包括：1）通过使用限制酶切割包含已知第一个核苷酸序列的DNA和与其相邻的第二个核苷酸序列，制备具有粘性末端的DNA片段; 2）用核苷酸类似物修饰DNA片段的3'末端以阻断DNA片段的进一步延伸; 3）将3'末端修饰的DNA片段的粘性末端与具有与其互补的内聚端序列的接头，接头或盒连接; 和4）使用已知的第一核苷酸序列特异性引物和接头，衔接子或盒序列特异性引物进行PCR。

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