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Macrophage specific targeting vector and recombinant cell constructed based on Ipr1 gene

A technology of targeting vectors and macrophages, applied in the direction of cells modified by introducing foreign genetic material, genetic engineering, plant genetic improvement, etc.

Active Publication Date: 2013-08-07
NORTHWEST A & F UNIV +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the current treatment of tuberculosis can only be treated with antibiotics, such as: streptomycin, kanamycin, etc.

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  • Macrophage specific targeting vector and recombinant cell constructed based on Ipr1 gene
  • Macrophage specific targeting vector and recombinant cell constructed based on Ipr1 gene
  • Macrophage specific targeting vector and recombinant cell constructed based on Ipr1 gene

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Embodiment Construction

[0042] The present invention first amplifies the MSR1 promoter and the homologous long arm and short arm from the bovine blood genome, and connects the Ipr1 gene and the above-mentioned fragments to the universal targeting vector pA2T to construct a macrophage-specific targeting vector pLMIS, and then uses The linearized pLMIS was introduced into neonatal bovine ear fibroblasts by electroporation, positive cell clones were obtained by G418 and GCV positive and negative screening, and identified by PCR and Southern blot, it was confirmed that the target gene Ipr1 was integrated into the genome of neonatal bovine ear fibroblasts. The detection results of the generated cloned cattle showed that the obtained cattle were transgenic cloned cattle with site-specific insertion of Ipr1, and could resist M.bovis infection.

[0043] The present invention will be further described in detail below in conjunction with the accompanying drawings and experiments, which are to explain rather tha...

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Abstract

The invention discloses a macrophage specific targeting vector and a recombinant cell constructed based on an Ipr1 gene. According to the invention, an MSR1 promoter is connected to a 5' terminal of the Ipr1 gene, such that the Ipr1 gene is specifically expressed in bovine macrophage with high efficiency; a recombinant gene LA-MSR1-Ipr1-SA is further constructed, such that a target gene is subject to site-directed integration, and is integrated between bovine 28-chromosome SP-A and MAT1A gene. An Ipr1 macrophage specific targeting vector pLMIS is constructed, and is transfected into newborn bovine ear fibroblast cells; through G418 and GCV positive and negative screening, transgenic newborn bovine ear fibroblast cell lines are constructed; the transgenic newborn bovine ear fibroblast celllines are adopted as nuclear donors, and are transplanted into bovine enucleated oocytes, such that transgenic cloned embryos are obtained; when the embryos are transplanted, five cattle with site-directed inserted Ipr1 gene is produced, and four of them survive. The transgenic cattle is resistant to M.bovis.

Description

technical field [0001] The invention belongs to the technical field of transgenic cloning animals, and relates to a macrophage heterosexual expression vector and recombinant cells constructed based on Ipr1 gene. Background technique [0002] Bovine tuberculosis, a zoonotic disease caused by M. bovis, is one of the most devastating bovine diseases in developing countries (Berrada and Barjas-Rojas, 1995). Mycobacterium bovis mainly infects cattle. Bovine tuberculosis is a zoonotic chronic infectious disease caused by Mycobacterium bovis. The diseased cattle are characterized by gradual emaciation and the formation of nodular granulation in tissues and organs. swelling and caseous necrosis. The cause of bovine tuberculosis is its easy infection and serious harm. Therefore, the World Organization for Animal Health lists it as a B-type infectious disease, and my country lists it as a second-class animal infection. Its spread affects the sustainable development of animal husband...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/85C12N15/12C12N5/10C12N15/877
Inventor 张涌何小宁权富生王勇胜
Owner NORTHWEST A & F UNIV
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