新城疫病毒F蛋白在昆虫细胞中的表达及其融细胞作用.pdfVIP

第 21 卷 6 期 中 国 病 毒 学 21 （6 ）：571-574 2006 年 11 月 VIROLOGICA SINICA November 2006 * 新城疫病毒 F 蛋白在昆虫细胞中的表达及其融细胞作用 ** 王桂军，秦爱建 ，邵红霞，崔平福，金文杰 （扬州大学江苏省动物预防医学重点实验室, 江苏扬州 225009 ） Expression and Characterization of the Fusion Protein of Newcastle Disease Virus Strain F48E8 in Insect Cells ** WANG Gui-jun ，QIN Ai-jian ，SHAO Hong-xia，CUI Ping-fu，JIN Wen-jie （Key Lab of Jiangsu Preventive Veterinary Medicine, Yangzhou University, Yangzhou 225009, China ） Abstract ：The fusion protein gene of strain F48E8 of Newcastle disease virus was amplified with RT-PCR, and cloned into the pGEM-T vector and named pGEM-NDF. Then, F gene was released from pGEM-NDF by digestion with BamHI and Xba I, and ligated to baculovirus transfer vector, pFastBac1, which was digested with BamHI and Xba I. The ligation yielded a recombinant plasmid, pFast-NDF. The results of sequencing and PCR showed that F gene had been cloned into baculovirus genome correctly. After transfecting sf9 cells with pFast-NDF, the recombinant baculovirus was recovered from the cells. The results of IFA and Western-blot assay indicated that F gene was expressed well in sf9 cells. Furthermore, the F protein was present on the cell membrane, and sf9 cells were fused at 96 hours post-infection, which suggested that F protein had the activity of cell fusion. Animal tests showed that the fusion protein expressed in insect cells induced neutralizing anti