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Fa-chao SUN Min TAN Yuan-chao ZHANG Yu-chao WANG Sheng-liang CAO Guo-fei DING Fang-yuan CONG Li-hong GUO Si-dang LIU Yi-hong XIAO 《农业科学学报》2019,18(7):1436-1442
To investigate the epizootic of swine influenza virus(SIV), 60 nasal swabs were collected from a clinical cases of pig farm in Tai'an City, Shandong Province of China in April 2017. SIV was isolated by inoculating into 10-day-old Special Pathogen Free embryonated eggs and the whole genome was sequenced. An H1N1 subtype SIV was isolated and designated as A/swine/Shandong/TA04/2017(H1N1). Phylogenetic analysis showed that apart from the polymerase A(PA) fragment belonging to the 2009 pandemic H1N1 branch, seven genome segments belonged to avian-like H1N1 influenza virus lineage. The cleavage site sequence of the hemagglutinin(HA) protein was PSIQSR↓G, which is a typical molecular biological characteristic. Five potential N-glycosylation sites(N14, N26, N277, N484 and N543) were found in the HA gene. To further investigate the epidemiology of SIV in this farm, the 995 serum samples were assessed with EAH1N1 2009 pandemic H1N1 and H3 N2 antigens. The results showed that the total positive rate was 65.43%. The positive rates of single virus infection detected by EAH1N1, 2009 pdmH1N1 and H3 N2 for serum HI(Hemagglutination inhibition) were 48.35, 30.85 and 7.47%, respectively. The results showed that SIV in Shandong Province has been reassorted in some segments and the SIV-positive rate was high on the SIV outbreak farm. These data provide evidence of an epizootic of SIV. 相似文献
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旨在建立鸡血液样品中多种喹诺酮类药物残留的超高效液相色谱—串联质谱方法。样品经乙腈提取,经Waters Acquity UPLC BEH C18色谱柱分离,以甲醇和0.1%甲酸水溶液为流动相进行梯度洗脱,电喷雾正离子(ESI+)模式电离,MRM扫描模式检测。经方法学验证,该方法的线性关系、回收率、精密度均符合要求。应用超高效液相色谱—串联质谱法建立了鸡血液样品中多种喹诺酮类药物同时检测的方法,实现了动物血液样品中12种喹诺酮类药物同时定性、定量分析。 相似文献
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参考 Genbank收录的 TGEV- Miller株的基因序列 ,自行设计合成 1对引物 (TGEVP5 /P6 ) ,对不同代次的 TGEV疫苗弱毒 STC3及种毒 、种毒 进行了 RT- PCR扩增 ,产物经琼脂糖凝胶电泳分析 ,均出现 1条大约 12 6 2 bp的目的条带 ,经 Eco R 酶切 ,都产生了 871bp和391bp左右的两个片段 ,与预期大小相符。将种毒 RT- PCR扩增目的条带回收纯化后克隆入PMD18- T载体中 ,转化宿主菌 DH5 α,挑选阳性克隆 (命名为 PTs) ,提取重组质粒 ,用 Hpa 、Eco R 对重组质粒进行酶切鉴定以及 PCR扩增 ,然后进行序列测定 ,并进行了序列分析 ,证实与国外标准毒株 Miller、Fs772 / 70、Purdue、TO14等有较高的同源性 相似文献
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抗霉菌素“120”是一种新的农用抗菌素,防治农作物上多种病害都有效。实验室试验证明;本剂与杀灭菌酯混用,对防治病害和虫害的效果没有不良影响,因此可以减少打药次数,有一定经济意义。 相似文献
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AIM: To investigate the effect of cGMP on voltage-gated potassium channel in pulmonary artery smooth muscle cells (PASMCs) from rats exposed to chronic hypoxia. METHODS: (1) Wistar rats were randomly divided into control group (group A) and chronic hypoxia group (group B). Then group B received hypoxia 8 hours per day for 4 consecutive weeks. (2) Single PASMC was obtained via acute enzyme separation method. (3) Conventional whole-cell patch clamp technique was used to record resting membrane potential (Em) and ion currents of voltage-gated potassium channel. The changes of ion currents of voltage-gated potassium channel before and after applying cGMP (1 mmol/L), an agonist of protein kinase G (PKG), and cGMP plus H-8 (1 mmol/L), an inhibitor of PKG were compared between two groups. RESULTS: The Em of group B were significantly lower than that of group A. The ion currents of voltage-gated potassium channel in group A and group B were all significantly inhibited by cGMP [control group: from (118.0±5.0) pA/pF to (89.9±16.5) pA/pF, n=6, P<0.05;chronic hypoxia group: from (81.0±5.0) pA/pF to (56.8±9.1) pA/pF, n=6, P<0.05]and these effects were reversed by H-8 [control group: from (119.2±10.3) pA/pF to (117.8±9.1) pA/pF, n=6, P>0.05;chronic hypoxia group: from (96.8±6.2) pA/pF to (98.0±2.2) pA/pF, n=6, P>0.05]. CONCLUSIONS: The currents of voltage-gated potassium channel was inhibited by chronic hypoxic. The inhibitory effect of cGMP on currents of voltage-gated potassium channel in PASMCs from both normal and chronic hypoxic rats may be probably through the phosphorylation of voltage-gated potassium channel. 相似文献
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表达H3N2亚型猪流感病毒HA基因重组伪狂犬病病毒的构建 总被引:4,自引:1,他引:4
将SV40启动子控制下的LacZ基因表达盒和CMV启动子控制下的H3N2亚型猪流感病毒(SIV H3N2)的HA基因插入到伪狂犬病病毒(PRV)通用转移载体pBdTK-Uni中,获得转移载体pLTK-HA。将该载体与PRV Bartha-K61株基因组DNA通过脂质体法共转染Vero细胞,经过10代蓝斑筛选、纯化和PCR鉴定获得了一株插入SIV HA基因的重组伪狂犬病病毒,命名为rPRV-HA。Western blotting和间接免疫荧光试验证实HA基因在重组病毒感染的细胞中获得了表达。用不同的细胞(PK-15、IBRS-2、Vero和鸡胚成纤维细胞)对该重组病毒与亲本病毒的增殖滴度和致细胞病变进行比较,未见显著差异,对第30代重组病毒的HA基因进行序列分析,表明该重组病毒遗传性状稳定。 相似文献