In order to reveal variation and revolution of NP genes of human avian H_5N_1 influenza virus strains,the NP gene of a human avian H_5N_1 influenza virus strain in Guangdong was sequenced and the global NP genes of st...In order to reveal variation and revolution of NP genes of human avian H_5N_1 influenza virus strains,the NP gene of a human avian H_5N_1 influenza virus strain in Guangdong was sequenced and the global NP genes of strains were retrieved.The sequences were analyzed by DNAStar 5.0,and the evolu- tionary speed was studied with reference to the epidemiological data.It was found that NP genes of 45 strains during 1997-2006 were homologically classified into three groups:strains in 1997-1998,strains in 2004-2005 and strains from 2003 to 2006.There were 35 substitutions in NPs in all strains accounting for a ratio of 7.03%(35/498).An additional glycoprotein domain(NGT_(430-432))was found in NP genes in the strains of 2003-2006,the mutation of N_(370)S in GD-01-06 resulted in occurrence of one more glyco- protein domain(NES_(368-370)).In the synonymous variation,K_s values in NP were 2.03×10^(-5)-2.55×10^(-5) Nt/d and K_a values in NP were 1.58×10(-6)-3.10×10^(-6) Nt/d.There didn′t exist obviously selective pres- sure.An additional glycoprotein domain in every strain of 2003-2006 and one more in strain GD-01-06 might change the antigenicity of human avian H_6N_1 influenza virus.The variation on human avian H_5N_1 influenza strains occurred frequently in the natural world,which would result in high probability of hu- man-human transmission along with the natural evolution of the virus.展开更多
为在昆虫细胞中表达鸭源新城疫病毒NP蛋白质,首先根据鸭源新城疫病毒NP基因序列设计引物,扩增出NP基因,将其克隆至杆状病毒表达载体p Fast Bac1,获得重组转移载体p Fast Bac-NP,将其转化到DH10Bac感受态细胞中,经抗性和蓝白斑筛选,获得...为在昆虫细胞中表达鸭源新城疫病毒NP蛋白质,首先根据鸭源新城疫病毒NP基因序列设计引物,扩增出NP基因,将其克隆至杆状病毒表达载体p Fast Bac1,获得重组转移载体p Fast Bac-NP,将其转化到DH10Bac感受态细胞中,经抗性和蓝白斑筛选,获得重组穿梭质粒r Bacmid-NP,在脂质体的介导下转染昆虫细胞Sf9,获得重组杆状病毒NP蛋白质。Western blot、间接免疫荧光试验结果均显示表达的重组蛋白质能够与全病毒阳性血清发生特异性反应,具有良好的反应原性。以上结果说明鸭源新城疫病毒NP蛋白质在昆虫细胞中获得了成功表达。展开更多
文摘In order to reveal variation and revolution of NP genes of human avian H_5N_1 influenza virus strains,the NP gene of a human avian H_5N_1 influenza virus strain in Guangdong was sequenced and the global NP genes of strains were retrieved.The sequences were analyzed by DNAStar 5.0,and the evolu- tionary speed was studied with reference to the epidemiological data.It was found that NP genes of 45 strains during 1997-2006 were homologically classified into three groups:strains in 1997-1998,strains in 2004-2005 and strains from 2003 to 2006.There were 35 substitutions in NPs in all strains accounting for a ratio of 7.03%(35/498).An additional glycoprotein domain(NGT_(430-432))was found in NP genes in the strains of 2003-2006,the mutation of N_(370)S in GD-01-06 resulted in occurrence of one more glyco- protein domain(NES_(368-370)).In the synonymous variation,K_s values in NP were 2.03×10^(-5)-2.55×10^(-5) Nt/d and K_a values in NP were 1.58×10(-6)-3.10×10^(-6) Nt/d.There didn′t exist obviously selective pres- sure.An additional glycoprotein domain in every strain of 2003-2006 and one more in strain GD-01-06 might change the antigenicity of human avian H_6N_1 influenza virus.The variation on human avian H_5N_1 influenza strains occurred frequently in the natural world,which would result in high probability of hu- man-human transmission along with the natural evolution of the virus.
文摘为在昆虫细胞中表达鸭源新城疫病毒NP蛋白质,首先根据鸭源新城疫病毒NP基因序列设计引物,扩增出NP基因,将其克隆至杆状病毒表达载体p Fast Bac1,获得重组转移载体p Fast Bac-NP,将其转化到DH10Bac感受态细胞中,经抗性和蓝白斑筛选,获得重组穿梭质粒r Bacmid-NP,在脂质体的介导下转染昆虫细胞Sf9,获得重组杆状病毒NP蛋白质。Western blot、间接免疫荧光试验结果均显示表达的重组蛋白质能够与全病毒阳性血清发生特异性反应,具有良好的反应原性。以上结果说明鸭源新城疫病毒NP蛋白质在昆虫细胞中获得了成功表达。
