摘要
目的探索小鼠β防御素2(mouseβ-defensin 2,mBD2)表达载体在狗肾传代细胞(Madin-Darby canine kidney,MDCK)中的表达和抗甲型流感病毒(influenza Avirus,IAV)活性。方法将真核表达载体pcDNA3.1/mBD2利用脂质体法转染MDCK细胞,G418筛选得到稳定转染的阳性细胞,用RT-PCR方法鉴定目的基因和免疫荧光法鉴定目的蛋白在转染细胞中的表达。用TCID50测定转染细胞的抗IAV活性。用pcDNA3.1/mBD2质粒免疫小鼠,观察死亡保护率。结果 pcDNA3.1/mBD2转染MDCK细胞后在细胞中稳定表达,转染细胞的TCID50比对照组的病毒效价降低近77.98倍;重组质粒免疫小鼠后,死亡保护率为55.55%。结论 mBD2能够在MDCK细胞中稳定表达,并且在体外及体内实验中显示出抗流感病毒活性,该结果为抗流感病毒制剂研究奠定了实验基础。
The objective of this paper is to study the expression of mBD2 in MDCK cell and its activity for against IAV.The established eukaryotic expressing plasmid pcDNA3.1/mBD2 was transfected into MDCK cell by PolyFect transfection Reagent.The positive clone with pcDNA3.1/mBD2 was screening with G418.Expression of target gene and peptide in transfected cells was detected with RT-PCR and the immunofluorescence assays,respectively.The activity of anti-IAV of transfected cells was measured with TCID50.Additionally,pcDNA3.1/mBD2 was injected into mice to observe its effect against IAV in vivo.The results showed that pcDNA3.1/mBD2 plasmid was transfected successfully and expressed stably in MDCK.The TCID50 on transfected cells decreased 77.98 times compared with control group,and it had distinct difference between the experiment and control groups（P〈0.05）.The protection rate for the mice immuned with pcDNA3.1/mBD2 after challenged with lethal IAV was 55.55%.These results suggested mBD2 has a clear activity for against IAV in vivo and in vitro,and it established a foundation for studying mBD2 against IAV.
出处
《中国人兽共患病学报》
CAS
CSCD
北大核心
2011年第9期792-795,共4页
Chinese Journal of Zoonoses
基金
国家自然科学基金资助项目(30671964)
