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当前位置: 首页 > 文章 > 口蹄疫病毒A型SAP突变毒株的构建及其生物学特性的测定 中国兽医科学 2014 (10) 1003-1007
Position: Home > Articles > Construction of foot-and-mouth disease virus type A with SAP mutation and detection of the mutation strain Chinese Veterinary Science 2014 (10) 1003-1007

口蹄疫病毒A型SAP突变毒株的构建及其生物学特性的测定

作  者：
杨洋;杨帆;杨波;郑海学
单  位：
中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室农业部畜禽病毒学重点开放实验室国家口蹄疫参考实验室
关键词：
A型口蹄疫病毒;反向遗传操作;SAP突变;病毒拯救
摘  要：
为了获得对牛和猪低致病性或无致病性的口蹄疫(FMD)疫苗毒株,提高疫苗毒株的生物安全性,选择口蹄疫病毒(FMDV)L蛋白酶编码基因SAP区域进行突变。利用反向遗传操作技术,构建FMDV SAP区域突变的A型病毒重组质粒prA-SAP-FMDV。利用脂质体将构建的重组质粒转染BHK21细胞,重组质粒能够致细胞病变(CPE)。连续传代后,经RT-PCR鉴定和间接免疫荧光试验等,SAP突变的重组病毒rA-SAP-FMDV被成功拯救。另外,比较测定了该重组毒与亲本毒rA-WT-FMDV对BHK21细胞和乳鼠的致病性;结果显示,rA-SAP-FMDV的TCID50/mL为10-7.17,LD50为10-6.5/0.2mL,与亲本毒株的生物学特性相似。这一突变SAP域的A型FMDV的获得为研发生物安全性高的A型FMD疫苗奠定了基础。
译  名：
Construction of foot-and-mouth disease virus type A with SAP mutation and detection of the mutation strain
作  者：
YANG Yang;YANG Fan;YANG Bo;ZHENG Hai-xue;State Key Laboratory of Veterinary Etiological Biology/Key Laboratory of Animal Virology,Ministry of Agriculture/National Foot-and-Mouth Disease Reference Laboratory/Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences;
关键词：
foot-and-mouth disease virus type A;;reverse genetics;;SAP mutation;;virus rescue
摘  要：
To obtain a new recombinant vaccine strain with higher biosecurity,SAP domain in leader proteinase of foot-and-mouth disease virus(FMDV)was mutated.SAP motif of L protein was selected and reverse genetics was used to obtain a FMDV type A recombinant plasmid named prA-SAP-FMDV.The prA-SAP-FMDV was transfected into BHK21 cells.The results of typical cytopathic effect(CPE),RT-PCR and indirect immunofluorescence assay showed that the recombinant virus,being named rA-SAP-FMDV,was rescued successfully.In addition,the pathogenicity of rA-SAP-FMDV was compared with that of the parental virus rA-WT-FMDV in BHK21 cells and suckling mouse.In result,the median tissue culture infection dose per mL(TCID50/mL)of rA-SAP-FMDV was 10-7.17,and its median lethal dose(LD50)was10-6.5/0.2mL.The results of TCID50/mL and LD50/0.2mL further showed that they were similar to those of rA-WT-FMDV on the biological property.The recombinant FMDV with SAP mutation will pave a way for study on type A FMDV strain with higher biosecurity.

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