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DNA表达载体直接注射哺乳动物肌肉组织后可转染肌肉细胞并表达基因产物。但这一方法具有组织局限性,除肌肉组织外,其他器官的组织很难通过这一方法将基因转移至细胞内并表达产物,本文的研究表明,甲状腺滤泡细胞具有肌肉相似的作用,可通过DNA直接注射法转基因。 本研究将携带氯霉素乙酰基转移酶(CAT)基因的表达载体(PCWV-CAT),和携带大肠杆菌β-半乳糖苷酶(β-gal)表达基因的PCMVnlacF,直接注射至兔甲状腺间隙组织和其他器官的组织中,3d后测定注射部位组织的CAT活性,并用X-Gal组织化学染色法鉴定DNA转染以及β-半乳糖苷酶的表达,发现在注射200μg剂量的DNA,3d后甲状腺滤泡细胞和肌肉中含有高表达活性的CAT,即在注射3d后,收集组织中单位质量的DNA或蛋白质中所含的CAT活性,肌肉中为80mUCAT/μgDNA或80mUCAT/μg蛋白质,甲状腺滤泡中含69mUCAT/μgDNA或83mUCAT/μg蛋白质;
DNA expression vectors can directly transfect mammalian muscle tissue to transfect muscle cells and express gene products. However, this method has the organizational limitations. In addition to muscle tissue, tissues of other organs are very difficult to transfer the gene into the cell by this method and express the product. The research in this paper shows that the thyroid follicular cells have similar muscle function, Transgenic by DNA direct injection. In this study, PCV-CAT carrying the chloramphenicol acetyltransferase (CAT) gene and PCMVnlacF carrying the gene encoding β-galactosidase (β-gal) of Escherichia coli were directly injected into the rabbit thyroid gland Tissue and other organs of the organization, the activity of CAT at the injection site was measured after 3 days and the DNA transfection and the expression of β-galactosidase were identified by X-Gal histochemical staining. It was found that after injecting 200 μg of DNA for 3 days Thyroid follicular cells and muscle contain highly expressed CAT, that is, after 3 days of injection, CAT activity per unit mass of DNA or protein in the tissue is collected, 80 mUCAT / μg DNA or 80 mUCAT / μg protein in muscle, and thyroid filtration Bubbles contain 69 mUCAT / μg DNA or 83 mUCAT / μg protein;