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Recombinant adenovirus of human hephaestin gene, preparation method thereof and application thereof

A ferroportin and auxiliary protein technology, applied in the field of genetic engineering, can solve problems such as difficulty in ensuring natural characteristics, instability, and low activity, and achieve the effects of treating neurodegenerative diseases, reducing toxicity, and high virus titer

Inactive Publication Date: 2010-11-03
THE HONG KONG POLYTECHNIC UNIV SHENZHEN RES INST
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Problems solved by technology

[0009] However, the use of exogenous protein therapy has disadvantages such as instability, low activity, high cost, and immunogenicity, and it is difficult to guarantee the natural characteristics of the secondary and advanced spatial structure of the protein when it exerts pharmacological effects. Express ferroportin auxiliary protein, increase the level of ferroportin auxiliary protein in vivo, regulate intracellular iron concentration, reduce the toxicity of iron to cells, and achieve the purpose of treating iron metabolism-related diseases such as neurodegenerative diseases

Method used

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  • Recombinant adenovirus of human hephaestin gene, preparation method thereof and application thereof
  • Recombinant adenovirus of human hephaestin gene, preparation method thereof and application thereof
  • Recombinant adenovirus of human hephaestin gene, preparation method thereof and application thereof

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Embodiment 1

[0066] Ferroportin Accessory Protein Gene Insertion Recombination

[0067] pTrack-CMV is a shuttle plasmid for recombinant adenovirus. We connected the two ends of the ferroportin auxiliary protein gene with Bgl II and Sal I restriction sites, and added a polyA tail as a termination signal region. The ferroportin auxiliary protein gene and the shuttle plasmid were digested with enzymes, and the two were ligated and transformed with T4 ligase. After screening, the correct clone was obtained and sent to the sequencing company for sequencing identification.

[0068] Ad-Heph structure: The recombinant human ferroportin auxiliary protein gene adenovirus (Ad-Heph) vector lacks the E1 region, and cannot be packaged into mature virus particles in ordinary cells, which is a defective virus. Human ferroportin auxiliary protein (hephaestin) gene was the target gene. We inserted the human channel-type ferroportin auxiliary protein gene into the genome of the virus, with double CMV as it...

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Abstract

The invention provides a recombinant adenovirus of a human hephaestin gene (Ad-Heph), which comprises the human hephaestin gene and an adenovirus vector. The invention also provides a preparation method and application of the recombinant adenovirus of the human hephaestin gene. In the invention, the human hephaestin gene is introduced into tissue cells of an organism by the defective adenovirus, the virus can autonomously express the hephaestin stably, safely and actively in the body of a patient, and thus, the hephaestin level in vivo is improved and the intracellular iron concentration is adjusted.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a recombinant human ferroportin auxiliary protein gene adenovirus carrying human ferroportin auxiliary protein (Hephaestin) gene capable of autonomous expression in vivo, its preparation method and application. Background technique [0002] As early as 1962, Grewal et al. discovered a new type of hereditary microcytic hypochromic anemia in radiation-induced mutant mice. Due to its association with the X chromosome, such mice are known as sla (sex-synthetic anemia) mice. Vulpe et al. (1999) confirmed the existence of defective genes on the X chromosome of sla mice through gene profiling studies. This gene is named Hephaestin (HP or Heph). The radiation hybridization map and the PCR technique of somatic cells confirmed that the human HP gene is located on the X chromosome. Syed et al. (2002) reported that the HP gene has an open reading frame through cD2NA cloning an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/01C12N15/85A61K48/00A61P3/00C12R1/93
Inventor 钱忠明葛啸虎柯亚
Owner THE HONG KONG POLYTECHNIC UNIV SHENZHEN RES INST
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