本发明公开了一种FMDV蛋白酶抑制剂的筛选方法和抑制效果评价方法,在GFP中插入FMDV的3C蛋白酶相关裂解位点的序列,形成改造后的荧光蛋白报告蛋白,在该荧光蛋白报告蛋白质粒上,克隆表达FMDV3C蛋白酶,可用于筛选蛋白酶抑制剂以及对蛋白酶抑制剂活性的评价,最终形成FMDV蛋白酶抑制剂筛选及药效评价平台。本发明能够避免使用FMDV...
Two 01 type FMDV isolates out of a total of 6 FMDV field isolates (including 01 Geshur) gave a positive PCR while two other 01 isolates and two ASIA isolates were detected with the RNA polymerase gene primers but not with the VP1 primers. Serial dilutions of the RNA used in each ...
重组质粒pUC57-MM-VP1、pET-28a表达载体和抗FMDV的阳性血清由河南省农业科学院动物免疫学重点实验室保存。核酸Marker DL2000、PrimerSTAR Max DNA聚合酶、T4 DNA连接酶、大肠杆菌感受态细胞JM109和BL21(DE3) 基因工程菌、伴侣蛋白质粒pTF16、感受态制备试剂盒购自Takara公司;胶回收试剂盒,质粒提取试剂盒等购自OMEGA公...
CSFVE2 FMDVVP1 5et al 2003 Diaz et al 2005 Wills et al 1997 Labaxque et al 2000 Gonin et al 1999 Meier et al 2003 El Kassar 1996Loemba et al 1996 Joo et al 1997 3PRRSVADE Fc Fc antibody dependent enhancement ADE Yoon 1996 PRRSV ADE PRRSV PRRSV PRRSV Yoon et al 1996 Yoon et al...
Fusion protein by the FMDV VP1 protein, a linker peptide and cytokine GM-CSF, whose amino acid residue sequence such as SEQ ID NO: 1 shown in Fig. 融合蛋白的编码基因由口蹄疫病毒VP1蛋白的编码基因,连接肽的编码基因和细胞因子GM-CSF的编码基因组成,其核苷酸序列如SEQ ID NO:3所示. Fusion protein...
1.2.2 FMDV病毒的鉴定和TCID50的测定 参照TaKaRa总核酸提取试剂盒说明书步骤,分别提取病毒RNA,反转录为cDNA,用Primer 5.0软件设计1对引物用以扩增VP1基因,序列分别为:VP1-P1:5′-CATGTCCTCTTGCCATCTGGTT-3′;VP1-P2:5′-AGATTTGTGAAAGTTAAGCCA-3′,PCR扩增条件为:94℃ 5 min;95℃ 30 s、55℃ 30 s、...
1988 年SVDV 的感染性cDNA 克隆成功构建,为了鉴定猪水疱病毒基因组中决定毒力的基因,Kanno T 等[11 ]在此基础上,从强毒株和弱毒株的cDNA 克隆制备了一系列重组病毒和定点突变病毒,最后证明基因组中决定毒力的基因在以下两个位点或其中的一个,一个是2842 位核苷酸,编码VP1 的132aa ,另一个是3355 位,编码2A...
1b). The resulting cDNAs were used as templates for RT-qPCR reactions on miRNA PCR array plates (Qiagen miScript Bovine miRNome) containing a series of primers unique to 169 characterized bovine miRNAs (Table 1) so as to identify miRNAs showing either up-regulation or down-regulation in the...
FMDV can enter cells using different pathways; the main pathway is by binding integrins via a conserved RGD located within the GH loop of VP1 [26, 27]. One subgroup within the integrin family is the αv integrin, which recognizes RGD as a binding motif on natural ligands. The αv ...
MicroRNAs play vital roles in regulating the battle between pathogens and host cells during viral challenging. MiR-4331 aggravates transmissible gastroenteritis virus (TGEV) -induced mitochondrial damage, also suppresses transcription of TGEV gene 7 via targeting cellular CDCA7. Otherwise, miR-4331-5p af...