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31.
试验旨在研究伪狂犬病病毒(PRV)在NF-κB家族p65基因敲除细胞系中的复制规律。利用慢病毒介导的CRISPR/Cas9基因定点修饰技术构建猪肺泡巨噬细胞(3D4/21)p65基因稳定敲除细胞系。通过构建p65-sgRNA重组质粒,转染至HEK293T/17细胞,收取慢病毒,感染3D4/21细胞后利用嘌呤霉素筛选获得多克隆细胞系,T7核酸酶检测敲除效率,再通过有限稀释法获得3D4/21-p65-/-的稳定细胞系。CCK-8试剂盒检测3D4/21细胞中敲除p65基因后对细胞增殖的影响;流式细胞术检测PRV-GFP感染3D4/21及3D4/21-p65-/-细胞后病毒增殖的差异;实时定量PCR检测PRV感染3D4/21及3D4/21-p65-/-细胞后PRV gB、TK基因mRNA表达水平及PRV感染细胞诱导的IL-1β和IL-6基因mRNA水平表达的变化;Western blotting检测PRV-QXX感染3D4/21及3D4/21-p65-/-细胞后PRV gB、gE蛋白的表达;滴度测定检测PRV-QXX感染3D4/21及3D4/21-p65-/-细胞后子代病毒滴度。结果表明,sgRNA2和sgRNA3的基因编辑效率较高,对其进行克隆化培养进而获得敲除p65基因的稳定表达细胞系;CCK-8试剂盒检测细胞活力表明,p65基因敲除对细胞活力无影响;流式细胞仪检测表明,同一时间点PRV-GFP在3D4/21-p65-/-中的增殖显著高于对照细胞;实时荧光定量PCR表明在3D4/21细胞中敲除p65基因促进了PRV gB、TK基因的mRNA表达水平,而抑制了IL-1β、IL-6基因的mRNA表达;Western blotting结果表明,在3D4/21细胞中敲除p65基因促进了PRV gB、gE蛋白的表达;滴度测定结果表明,同一时间点PRV-QXX在3D4/21-p65-/-细胞中子代病毒的复制显著高于对照细胞。以上结果均表明,p65基因敲除可促进PRV在3D4/21细胞中复制。 相似文献
32.
Influence of four soil maintenance practices on Collembola communities in a Mediterranean vineyard 总被引:1,自引:0,他引:1
Anglique Renaud Nicole Poinsot-Balaguer Jrme Cortet Jean Le Petit 《Pedobiologia》2004,48(5-6):623-630
We studied the influence of four soil maintenance practices on Collembola communities in the soil of a Mediterranean vineyard: (a) postemergence herbicide with glyphosate; (b) postemergence and pre-emergence herbicides with glyphosate, terbuthylazine, diuron and oryzalin; (c) natural flora and (d) tillage to a depth of 10–15 cm. Total Collembola abundance, species diversity and species richness significantly varied between the four practices. Notably, the practice using postemergence and pre-emergence herbicides had significantly lower values. Identification of Collembola at species level allowed an interspecies comparison and revealed significant differences for the most common species between the four practices, with each practice being characterized by a different set of species. None of the species were found to be significantly more abundant in the plots treated with postemergence and pre-emergence herbicides. 相似文献
33.
GUO Yuan LIU Zong-zheng LIU Le MENG Fan-hua ZHANG Yan-ru ZHOU Huan-min LIU Chun-xia 《中国畜牧兽医》2016,43(11):3011-3018
In order to obtain the cloned sheep using bone marrow mesenchymal stem cells(BMSCs)as the donor cells,the BMSCs of sheep were chosen and reconstructed embryos were built to transfer.10 published microsatellite markers were chosen,and the DNA samples from clone sheep,donor cells and surrogate ewes were amplified,and the relationship of father-son(RCP)was analyzed using the Quantity One for genotyping.The results showed that the reconstructed embryos were successfully built for electric fusion using sheep BMSCs as nuclear donor,and making nuclear transplantation into enucleated mature oocytes of which the fusion rate was 80.62%.20 surrogate ewe were chosen to be implanted with the reconstructed embryos at morula stage by implant surgery,and 5 lambs were born and only 3 were survived.The genotype of cloned sheep was in line with the dornor cell and the RCP were more than 99.999%.In conclusion,the first clone sheep were obtained successfully by using BMSCs as a nuclear donor in this experiment. 相似文献
34.
The sustainability of self-governingirrigation schemes is currently underpressure in many countries as publicfinancial support is decreasing.Furthermore, growing global concernregarding water scarcity means thatefficient water use is essential.Theoretically the choice and implementationof a water-pricing system should play acentral role in achieving this objective,both by recovering water costs and byencouraging farmers to adopt more efficientbehaviour. In the Senegal River Delta newlyestablished water users associations (WUAs)have chosen low water charges, which areaffordable for the majority of farmers butwhich underestimate long-term maintenancecosts. Combined with the difficulty ofcollecting and managing farmers' fees, thischoice has drawn them into a vicious circleleading to scheme deterioration and poorwater service. New alternatives have beendiscussed with them using a simulation toolthat takes into account both the watercosts and the farmers' incomes. Comparedwith the present fixed water charge basedonly on cultivated land, a two-part optionbased both on equipped and irrigated areasensures the recovery of fixed expenses suchas maintenance, while encouraging farmersto intensify their farming systems byadopting double cropping. Neverthelesschoosing a relevant water charge does notensure by itself the sustainability of anirrigation scheme. Combined supportproviding WUAS and farmers with efficientadvice regarding management andorganisational skills should alsocontribute significantly to achieve thisprime objective. 相似文献
35.
目的 观察淫羊藿对肾阳虚雄性SD大鼠肾脏骨形态发生蛋白-7(BMP-7)表达的影响.方法 3月龄雄性SD大鼠24只,随机分为正常组、肾阳虚模型组、淫羊藿组,灌胃给药90d后,RT-PCR检测肾脏BMP-7 mRNA表达,免疫组化检测肾脏BMP-7的表达.结果 正常组和淫羊藿组BMP-7mRNA和蛋白显著高于肾阳虚模型组(灰度比值:0.990±0.103和1.054:t:0.097vs0.619±0.106,P<0.01;免疫组化:0、1、0、4和0、0、0、4vs1、2、2、0,P<0.01).结论 淫羊藿可提高肾脏中BMP-7的表达而保护肾脏. 相似文献
36.
针对南方稻田土壤酸化严重,导致养分流失有毒重金属活化,严重影响稻米质量安全的重大现实问题。以水稻秸秆和谷壳等农业废弃物为原料制备生物炭(分别记为RSC和RHC),研究不同原料生物炭对酸化土壤改良及其对重金属有效性的影响。设置3个生物炭用量(0,20,50 g/kg,分别记为CK、C1、C2),4种土壤酸化水平(pH 4.01,4.25,4.33,4.58,分别记为L1、L2、L3、L4),生物炭与重金属污染土壤共同培养60天后测定土壤pH、全氮、有机质、有效磷、速效钾和有效态Cu、Cd含量。结果表明:RSC对酸化土壤pH的改良效果明显优于RHC,且施炭量越高提高幅度越大,RSC的C2处理使4种酸度水平的土壤pH分别提高了0.68,0.97,1.29,1.71个单位。2种生物炭均能提高土壤的全氮、有效磷、速效钾和有机质含量,其中各施炭处理有机质显著提高,尤以速效钾的增幅最为显著,RSC对4种养分的提高均优于RHC。RHC对土壤有效态Cu含量无显著影响;RSC的C2较C1处理更能降低土壤中有效态Cu含量,使4种酸度水平的土壤分别降低了13.62%,6.57%,4.36%,7.88%。RHC处理的L3、L4土壤中有效态Cd含量显著降低,最大分别降低了13.79%,19.23%。RSC使4种酸度土壤有效态Cd含量最大分别降低了20.00%,25.81%,20.69%,19.23%。相关分析表明,土壤pH与有效态重金属含量呈显著负相关关系。水稻秸秆炭用于改良酸化土壤、降低重金属Cu和Cd有效性的效果更佳,且降低污染土壤中Cd的有效性较Cu好;生物炭对酸化程度越低的土壤pH和有效磷含量的提高以及有效态Cd含量的降低效果较好,而有效态Cu含量的降低效果则在酸化程度越高的土壤中表现更佳;土壤pH是生物炭调控重金属Cu、Cd有效性的主要影响因素。 相似文献
37.
杉木林下套种不同树种对土壤改良效应初探 总被引:5,自引:0,他引:5
为探讨复层异龄林营建初期对土壤物理结构和养分状况的影响,研究了浙西地区杉木林下套种不同树种土壤理化特征的变化。结果表明林下套种树种后林地土壤物理结构均得到不同程度的改善,土壤养分状况进一步好转。并运用多因素主成分分析方法,评价了不同复层林模式土壤养分含量规律,并将土壤养分状况的综合优势作了排序。其结果显示杉木林下套植的12种植物在种植初期对土壤养分改良效果较好的为麻栎、毛红椿、南天竺、十大功劳、红豆杉、椤木石楠和蚊母,其中麻栎和毛红椿改良效果最佳。 相似文献
38.
Residue quality has been shown to influence soil water-stable aggregation (WSA) during crop residue decomposition, but there is still little information about its interactive effect with soil mineral N availability. The aim of this study was to determine the effect of soil mineral N on WSA during the decomposition of two high-C/N crop residues (wheat straw with C/N = 125.6 and miscanthus straw with C/N = 311.3). The two crop residues were combined with three mineral N addition rates (0, 60, and 120 mg N kg−1 dry soil). Respiration, soil mineral N content, and WSA (expressed as mean-weight diameter, MWD) were measured on several dates during a 56-d incubation. The effect of decomposing crop residues on WSA followed two phases. (i) Between 0 and 7 d, the increase in WSA was related to intrinsic residue quality with higher decomposability of the wheat straw resulting in higher WSA. (ii) Thereafter, and until the end of the experiment, mineral N addition rates had a predominant but negative influence on WSA. In this second phase, the average MWD of residue-treated soils was 0.92, 0.55, and 0.44 mm for the 0, 60 and 120 mg N kg−1 dry soil addition rates, respectively. Mineral N addition which did result in higher crop residue decomposition did not lead to higher WSA. WSA during crop residue decomposition is therefore not simply positively related to the induced microbial activity, and changes in microbial community composition with differential effects on WSA must be involved. The impact of high-C/N crop residues inputs on WSA, initially assumed to be low, could actually be strong and long-lasting in situations with low soil mineral N content. 相似文献
39.
通过田间试验方法,研究了在铅锌矿废水污染的土壤上施用石灰1 a后,继续施用石灰和石灰后效处理对后续第1、2季低累积玉米(Zea mays)的产量以及重金属Cd、Pb、Zn和Cu含量的影响,并分析了土壤pH、土壤DTPA提取态重金属含量和土壤重金属全量的变化。结果表明,连续施用石灰和石灰后效均显著提高玉米产量,其中连续施用石灰处理效果最佳,第1季籽粒产量是对照(无石灰)的6倍,第2季是对照的3.8倍。与对照相比,连续施用石灰处理显著降低了2季玉米籽粒Cd、Pb、Zn和Cu含量,石灰后效只能降低第2季玉米茎叶Cd、Pb和Cu含量,而籽粒Cd、Pb含量与对照相比略有升高,说明石灰后效能维持一年半左右。对照处理土壤Cd和Zn全量显著低于石灰处理,可能是土壤中Cd和Zn随着雨水的淋洗向下层迁移造成的。施用石灰可防止Cd和Zn对地下水的污染。 相似文献
40.