动物造模,HIV病毒,转基因动物模型 z-bio 2022-01-10 856

　　(1) Copy method

　　① The HW-1 long terminal repeat (LTR) is related to the indicator gene of transgenic mice, that is, the LTTR gene is related to the indicator gene-chloramphenicol acetyltransferase (CAT). Already introduced. ②The tat gene sequence of HW-1tat transgenic mice, namely ARV-2 (AIDS-related retrovirus strain 2) virus, is connected to HIV-1 LTR and introduced into the PML plasmid. The tat sequence is 179 bases. It encodes the 8th-66th amino acid. These fragments were then injected into the fertilized egg cells of CD-1 male mice that were crossed with C57BL/6×DBA male mice. Newborn mice were tested by Southern blotting, and some mice expressed sequences on the skin. ③HIV full-sequence proviral DNA transgenic mice, that is, transgenic DNA transgenic mice established after the establishment of HIV-1 full-sequence disease. The T and t antigen genes of SV40 are complete virus-gene mice.

　　(2) Model characteristics

　　① HIV-1 long terminal repeat (LTR) and indicator genes are related to transgenic mice, eyes, heart, thymus and tail. When co-cultured with pHA or cytokines, peripheral blood lymphocytes and monocytes can also express CAT activity, and CAT activity is the highest in skin Langerhans cells. Immunocytochemical staining technique is used to detect the LTR transcriptional activity of HIV-1 in the cerebral cortex, basal ganglia and cerebellar glial or neuronal cells. This transgenic mouse expresses the HIV-1 LTR gene in multiple organs and tissues, but only expresses this gene and does not cause the disease corresponding to the mouse. ②The skin lesions of the male mice of HIV-1tat transgenic mice showed the proliferation and development of spindle cells, while the female mice had skin lesions or tumors, and the incidence of liver cancer was high. At this time, the growth factors released from the skin mediated by the tat gene indirectly affect the development of tumors. Also found out. In this transgenic mouse model, a large proportion of animals develop human-like Kaposi's sarcoma on the skin. This may be caused by HIV-1, especially the tat gene product.

　　③In HIV full-sequence proviral DNA transgenic mice, it has been confirmed that a complete copy of the virus gene is expressed in the mouse, but not all mice show signs of gene expression or disease. The F1 offspring produced by sexual reproduction of female mice and normal male mice expressing the complete sequence gene grow very slowly. Lymphocytes infiltrate around the blood vessels in the lungs, the epidermis is the tail, ears and nose, infiltrated legs, etc. .. Hyperplasia and lymphocytosis and thymic atrophy. However, the selective destruction of T lymphocytes and the phenotype of human AIDS-like diseases were not observed in F1 offspring. HIV-1 virus particles can be isolated from the affected F1 progeny, and have the ability to infect human CD4-positive cells, but cannot infect mouse fibroblasts. (3) Comparative medicine HIV-1 has been confirmed as the pathogen of AIDS. The relationship between HIV-1 infection and the AIDS outbreak remains unknown. Establishing an animal model of AIDS is an important tool for studying immunodeficiency caused by HIV-1 and determining the efficacy of vaccines or the therapeutic effects of antiviral drugs. So far, many animals (such as chimpanzees, gibbons, rhesus monkeys, rabbits and cats) have been tested in animal models of HIV-1 infection, but due to the source of certain large animals and economic reasons, HIV-1 is still Unsatisfactory reports on animal models. As an animal model, mice have received more and more attention from scholars all over the world. However, it cannot be used to study HIV-1 because mice lack CD4 receptors. Recent years. The successful research on genetically modified mice and severe combined immunodeficiency mice (SCID) makes it possible to use mice in animal models of HIV-1, ADS antiviral therapy and vaccine efficacy.