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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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CHENG Biao FU Xiao-bing SHENG Zhi-yong SUN Tong-zhu SUN Xiao-qing LI Jian-fu 《园艺学报》2002,18(11):1333-1336
AIM: To examine the expression of alpha-smooth muscle actin in scar tissue, and observe the phenomenon of apoptosis and its involvement in the process of pathological scarring and the presence of myofibroblasts or absence of cell in the dermis. To investigate the potential role of reparative cell apoptosis in hyperplastic scar formation. METHODS: The samples of scar were obtained from post-burn patients undergoing plastic operation in our burn unit recently, and the samples of control came from skin donor site of the same patient correspondingly. TUNEL assays were performed to evaluate the number of apoptotic cells in scar versus normal skin. In situ hybridization and immunohistochemistry staining technique were employed to determine the expression of different dermis cells markers in scar tissue and normal skin. RESULTS: There existed evident difference in apoptotic cells in the dermis between scars tissue and normal human skin. The expression positive cells were much more in hyperplastic scars than that in normal human skin; the apoptotic cells of proliferative stage were slight more than that of mature stage. However, in proliferative stage, the number of apoptotic cells was higher for the combination of hyperplastic scar than normally healed flat scars. But in mature stage, no obviously difference was detected between hyperplastic scar and normally healed flat scar. The monoclonal anti-α smooth muscle actin (ASMA) expression was significantly stronger in proliferative stage than that of mature stage. CONCLUSIONS: With reconstitution of dermal tissue, myofibroblasts containing alpha-SM actin disappear under normal wound healing, probably as a result of apoptosis. The myofibroblast play a critical role in wound closure and in the pathologic sequelae of healing. 相似文献
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枯草芽孢杆菌B—916防治水稻纹枯病的田间试验 总被引:35,自引:4,他引:35
1992~1994年在田间小区试验比较两个拮抗细菌(B-916、P-91)和井岗霉素对水稻纹枯病的防治效果。1994~1995年在江苏省3个基点上进行了B-916防治纹枯病的大面积示范试验。B-916发酵液在田间对水稻纹枯病的为害有较强的控制能力,发酵含菌量为2.54×1011~2.54×1012cfu/mL时,用量3.75L/hm2的防治效果为50.0%~81.0%。 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