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| 携带IL-2/NK4双基因减毒沙门氏菌TPIN的遗传稳定性研究 |
| Heredity Stability of Attenuated Salmonella TPIN carrying IL-2/NK4 Double Gene |
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| DOI：doi:10.3969/j.issn.1005-7021.2020.03.006 |
| 中文关键词: 基因治疗 减毒沙门氏菌 遗传稳定性 IL-2基因 NK4基因 |
| 英文关键词: gene therapy attenuated Salmonella heredity stability IL-2 gene NK4 gene |
| 基金项目:甘肃省重点研发计划项目（17YF1FA135） |
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| 中文摘要: |
| 对携带IL-2/NK4双基因减毒沙门氏菌TPIN的遗传稳定性进行研究。将TPIN连续传40代，取10代、20代、30代、40代菌落进行鉴定，包括菌落和菌体的形态、质粒的纯度和浓度、目的基因片段、双酶切结果及转染人肝癌细胞（HepG2）后目的基因体外表达活性。结果表明，每10代的菌落和菌体形态一致，其所含质粒的纯度和浓度在传代过程中无明显变化，PCR扩增的目的基因条带大小一致，双酶切鉴定结果正确，转染HepG2细胞后，每代间所表达目的蛋白浓度无明显差异。TPIN作为消化道肿瘤的生物治疗药物，在遗传方面有较好的稳定性，可进一步进行大量生产。 |
| 英文摘要: |
| The heredity stability of Salmonella TPIN carrying the IL-2/NK4 gene attenuated was studied. The TPIN was subcultured for 40 generations in succession. The colonies of 10 generations, 20 generations, 30 generations and 40 generations of TPIN were taken and identified, including the morphology of colony and cell, the purity and concentration of plasmid, the target gene fragment, the results of double enzyme digestion and the expression of the target gene after transfection into human hepatocellular carcinoma cell line (HepG2) in vitro. The results showed that the colonies and the morphology of the colonies of each 10 generations were consistent, the purity and concentration of plasmid had not changed significantly during the process of going down the generations. The size of the target gene bands amplified by PCR was consistent, and the results of double enzyme digestion were correct. There was no significant difference in the concentration of the target protein expressed among each generations after transfection of HepG2 cells. Therefore, it could be concluded that as a bio-therapeutic drug for digestive tract tumors, TPIN has comparatively good genetic stability and can be further produced in large quantities. |
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