• 专利附录
  • 专利说明
  • 权利要求
  • 类似技术
  • 同族专利
  • 引用文献
  • 法律状态
  • 优先权
    • 专利摘要:
    Gene construction and its use in the treatment of cardiac fibrosis. The present invention relates to a novel gene construct comprising a) a dna sequence encoding the prommp2 protein, b) a dna sequence encoding the MHC-β promoter, operably linked to sequence a), and c) a protein expression and secretion vector operatively linked to sequences a) and b). Also, a pharmaceutical composition, comprising said gene construction, is contemplated for its use in the treatment of cardiac fibrosis. Finally, the method for obtaining the gene construct of the invention is contemplated. (Machine-translation by Google Translate, not legally binding)
    公开号:ES2620702A1
    申请号:ES201700440
    申请日:2017-03-29
    公开日:2017-06-29
    发明作者:Ana Victoria VILLAR RAMOS
    申请人:Universidad de Cantabria；
    IPC主号:
    专利说明:

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    insertion of molecular DNA into a commercial gene expression vector (Sambrook et al, 1989).
    In general, the method for obtaining the gene construct of the invention (hereinafter the method of the invention) comprises the following steps:
    Linearize protein expression and secretion vector
    Enter the two DNA sequences a) and b) into the linearized vector (DNA sequence encoding the proMMP2 protein and DNA sequence encoding the MHCβ promoter) and
    Obtaining the gift "promoter of MHCβ-vector-proMMP2", or gene construct of the invention.
    In an intended embodiment, the protein expression and secretion vector employed in the method of the invention is the pSecTag2A vector.
    The method of obtaining this gene construct (MHCβ-pSecTag2A-proMMP2 promoter) comprises the following steps:
    i. Linearize the expression vector pSecTag2A in its region of multiple cloning sites (MCS) using restriction enzymes;
    ii. Introduce the proMMP2 (a) DNA sequence into the MCS region of the pSecTag2A (c) vector by using DNA ligase for the union of the two DNA fragments with complementary ends.
    iii. Obtaining the proMMP2-pSecTag2A clan;
    iv. Linearize the proMMP2-pSecTag2A clan and delete the CMV promoter region (cytomegalovirus) through the use of restriction enzymes;
    v Enter the DNA sequence of the MHCβ promoter (b) into the CMV promoter region of the vector previously deleted in iv) using DNA ligase; Y
    saw. Obtaining the clan "promoter of MHCβ-pSecTag2A-proMMP2", which constitutes a unique and intact DNA molecule.
    In a preferred embodiment of the method of the invention, in step vi), the obtained clan (MHCβ-pSecTag2A-proMMP2 promoter) has the sequence shown in SEQ ID NO
    Four.
    In another main aspect of the invention an isolated cell transfected with the gene construct of the invention is contemplated, where the vector allows the expression of the MMP2 protein and where said protein is secreted extracellularly.
    In particular embodiments of the invention, the cell is a cardiomyocyte.
    The transfection can be extended to other specific cell types that also possess the appropriate cellular machinery for MHCβ synthesis such as fibroblasts, osteoblasts, etc. Transfection of the proMMP2 gene can be carried out by
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    权利要求:
    Claims (1)
    [1]
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    类似技术:
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    同族专利:
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    ES2620702B2|2017-12-18|
    引用文献:
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    WO1996021416A2|1994-12-30|1996-07-18|Chiron Corporation|Methods and compositions for treatment of solid tumors in vivo|
    WO2001090326A2|2000-05-22|2001-11-29|Pharmacia & Upjohn Company|Novel matrix metalloproteinases|
    EP3081081A1|2013-12-12|2016-10-19|Mie University|Transgenic non-human mammal expressing human mmp2|
    法律状态:
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    申请号 | 申请日 | 专利标题
    ES201700440A|ES2620702B2|2017-03-29|2017-03-29|Gene construction and its use in the treatment of cardiac fibrosis|ES201700440A| ES2620702B2|2017-03-29|2017-03-29|Gene construction and its use in the treatment of cardiac fibrosis|
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