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991.
新城疫病毒F、NP、M和HN基因在昆虫细胞内的共表达 总被引:3,自引:0,他引:3
为构建杆状病毒转移载体,通过PCR的方法将F48E9株新城疫病毒F基因上的StuⅠ位点和NP基因上的XbaⅠ位点进行突变,扩增出全长的F、NP以及F48E9株新城疫病毒M和HN基因片段,将其克隆到pMD18-T载体,再将F、NP、M和HN基因依次亚克隆到杆状病毒转移载体pAeAB4上,F基因和M基因均在p10启动子的操控之下,NP基因和HN基因构建到两个polyhedrin启动子下游,构建重组杆状病毒转移载体pAeAB4-F-NP-M-HN。pAeAB4-F-NP-M-HN与线性化的杆状病毒DNA共转染昆虫细胞Sf9,获得重组杆状病毒rBae-F-NP-M-HN。重组杆状病毒感染Sf9细胞,72h后收集细胞和培养上清。Western blot分析显示:M、NP、F和HN蛋白在培养上清中得到了共表达,大小与预期结果一致;感染细胞内只检测到了HN蛋白的表达。这表明M、NP、F和HN蛋白在昆虫细胞内共表达可以自我装配成病毒样颗粒,并且以出芽的方式释放到培养基中。该重组杆状病毒的获得为研究新城疫病毒各结构蛋白之间的相互作用和确定病毒粒子出芽的驱动力等方面奠定了基础。 相似文献
992.
表达鸡马立克氏病病毒gB基因重组鸡痘病毒的遗传稳定性及生物安全性评价 总被引:1,自引:0,他引:1
为了评价表达鸡马立克氏病病毒gB基因重组鸡痘病毒(rFPV-gB/R)的遗传稳定性,我们将纯化后的重组病毒在CEF单层上连续传30代,引起细胞病变的速度和形态均未发生明显变化;覆盖含X-Gal的琼脂引起的空斑均为蓝色;间接免疫荧光实验证明rFPV-gB/R中的gB基因始终能稳定表达;序列测定结果表明,重组病毒在细胞上连续传30代、在SPF鸡上连续传5代后,gB基因序列没有发生任何变化;以0、10、20、30代重组病毒制成冻干疫苗进行的实验室免疫效力实验表明,细胞连续传代后rFPV-gB/R仍然保持了原有的免疫原性。可见重组鸡痘病毒gB基因的结构和免疫原性都是高度稳定的。为了评价rFPV-gB/R的生物安全性,我们将rFPV-gB/R通过SPF鸡连续传5代,检测病毒在鸡体的存在部位及其消长、生长繁殖性能和毒力变化;将rFPV-gB/R免疫鸡与未免疫鸡同笼饲养,攻击FPV-102E6强毒,以检测rFPV-gB/R感染鸡的接触传染性。结果显示,rFPV-gB/R在鸡体的存在时间大约为7d,在体内仅存在于接种部位;鸡体传代后痘病毒毒力有一定程度下降,gB基因核苷酸序列未发生任何变化;同居未免疫SPF鸡在痘病毒强毒攻击后全部发痘,可见重组病毒免疫鸡没有接触传染性,能在鸡体内稳定地传代,rFPV-gB/R具有高度的生物安全性。 相似文献
993.
研究了头孢噻呋对实验性感染鸡白痢沙门氏菌的防治效果。结果显示,头孢噻呋体内疗效良好,治疗组与预防组的治愈率、有效率、相对增重率均不同程度高于感染对照组(P〈0.05或P〈0.01);头孢噻呋的治疗效果高剂量组与中剂量组无显著差异(P〉0.05),高剂量组与低剂量组差异显著(P〈0.05);头孢噻呋的中剂量组与头孢曲松、氨苄西林组的疗效相比均差异显著(P〈0.05或P〈0.01)。实验结果表明,头孢噻呋按0.1mg/只的剂量肌肉注射,每日一次,可对鸡白痢沙门氏菌病起到有效的防治效果。 相似文献
994.
995.
AIM: To study the protective effects of bone marrow-derived mesenchymal stem cells on damaged dopaminergic neurons induced by 1-methyl-4-phenylpytidinium(MPP+).METHODS: The parkinson disease(PD) models were established in newborn rats. Bone marrow mesenchymal stem cells(MSCs) obtained from adult bone marrow were cultured, isolated and purified. MSCs were co-cultured with brain slice and the immunohistochemical technique, electron microscopy, propidium iodide staining were used to observe the changes of neurons. RESULTS: In the MPP+ treatment group, the neurites grew slowly and sparsely, dead cells were found in all regions. In the co-culture group, the neuritis grew densely, only a few cells were dead, the number of tyrosine hydroxylase(TH)-stained neurons increased and the structure of organellae was normal. CONCLUSION: MSCs may protect dopaminergic neurons against damage induced by MPP+. These results provide some data for cell transplantation therapy to Parkinsons disease. 相似文献
996.
997.
Yuuri Hirooka Takao Kobayashi Jun Takeuchi Tsuyoshi Ono Yasunori Ono Keiko T. Natsuaki 《Journal of General Plant Pathology》2007,73(5):330-335
A ring spot disease of Aloe vera was found on leaves of potted seedlings of Aloe vera in Hachijojima and Chichijima Islands, Tokyo. From tissue of ring spot lesions, a fungus producing Fusarium-type conidia was consistently isolated. After 1 month, reddish perithecia of nectriaceous fungus had formed on the colonies
of this isolate on PDA. These nectriaceous and Fusarium fungi were identified as Haematonectria haematococca and Fusarium sp., respectively. From a single ascospore isolation, the former was confirmed to be the teleomorph of the Fusarium sp. Typical ring spot lesions were reproduced by artificial inoculations using single ascospore and single conidium isolates.
Inoculations of five species of genus Aloe revealed that they were highly susceptible except for A. arborescens. This is the first report of a disease on Aloe caused by H. haematococca (anamorph: Fusarium sp.) in Japan, and it was named aloe ring spot. 相似文献
998.
Taketo Ashizawa Masashi Sasahara Atsushi Ohba Takeshi Hori Kouji Ishikawa Yukio Sasaki Tomohisa Kuroda Ryoei Harasawa Kaoru Zenbayashi-Sawata Shinzo Koizumi 《Journal of General Plant Pathology》2007,73(1):15-21
Leaf blast suppression in multilines was evaluated based on the number of susceptible lesions observed in a pure stand of
susceptible rice cultivar Sasanishiki, and in 1 : 1 and 1 : 3 mixtures of Sasanishiki and a resistant near-isogenic line,
Sasanishiki BL4 or BL7, from 1998 to 2001. The number of lesions first observed in fields in the 1 : 1 and 1 : 3 mixtures
were close to theoretical numbers calculated using the number of lesions observed in the pure stands and the ratios of the
susceptible Sasanishiki in the mixtures. The ratio of the number of lesions in the 1 : 1 and 1 : 3 mixtures to the number
in the pure stand was 0.29 and 0.09, respectively. The relationship between these ratios and the ratios of susceptible Sasanishiki
in mixtures was defined in an equation to estimate the degree of leaf blast suppression. Validation studies for the ratios
of the number of lesions in the 1 : 1 and 1 : 3 mixtures to the number in the pure stand were conducted in two different locations
and showed that the ratios are almost acceptable. The calculated autoinfection to alloinfection ratio was 1.3 and 1.4 in the
1 : 1 and 1 : 3 mixtures, respectively, suggesting that the calculated ratio will affect the degree of leaf blast suppression.
Thus, predictors were obtained to estimate leaf blast suppression for effective blast control in multilines. 相似文献
999.
Alessandro Zanzotto Federica Autiero Diego Bellotto Gianluca Dal Cortivo Gianluca Lucchetta Michele Borgo 《European journal of plant pathology / European Foundation for Plant Pathology》2007,119(2):183-192
During a 6-year study, grapevine propagation materials and young grapevines were analysed to evaluate the presence of internal
wood discolouration and the occurrence of fungal species involved in Petri disease. The intensity of wood discolouration increased
with the ageing of the plants. The maximum incidence of dark streaks was observed in the rootstock while necrosis originating
from buds or nodes were notably present in the trunk and cordon of older vines. In contrast, the highest levels of brown-red
halo symptoms, defined as discoloured areas around the pith, were recorded in the early growth stages. Phaeoacremonium spp. and Phaeomoniella chlamydospora were usually isolated from the rooted-grafts and the 3-year old plants, respectively. The number of infected grapevines increased
with age. Most of the P. chlamydospora strains were isolated from dark streaks or dots, while Phaeoacremonium spp. were detected in brown-red halo symptoms and other symptomatic or asymptomatic wood. The greatest incidence of the two
fungal taxa was recorded in the lower parts of the grapevine, including the roots and rootstock. 相似文献
1000.
Control of black foot disease in grapevine nurseries 总被引:1,自引:0,他引:1
Black foot disease of grapevines is a decline and dieback disease caused by a soilborne pathogen complex including Cylindrocarpon liriodendri , C. macrodidymum , Campylocarpon fasciculare and Campyl. pseudofasciculare . These pathogens cause primary infections of roots and basal ends of grafted cuttings in nursery soils. Thirteen fungicides were screened in vitro for mycelial inhibition of these pathogens. Prochloraz manganese chloride, benomyl, flusilazole and imazalil were the most effective fungicides tested, and were subsequently included in semi-commercial field trials. Basal ends of grafted cuttings were dipped in various chemical and biological treatments prior to planting in open-rooted nurseries. Black foot pathogens were not isolated from grafted cuttings prior to planting. Additional treatments involved soil amendments with Trichoderma formulations and hot water treatment of dormant nursery grapevines. Field trials were evaluated after eight months. Isolations from uprooted plants revealed low levels of black foot pathogens in the roots of untreated control plants, and significantly higher levels in basal ends of rootstocks. The incidence of black foot pathogens, as well as that of Petri disease pathogens, was not significantly and/or consistently reduced by the majority of chemical or biological treatments. However, these pathogens were not isolated from uprooted plants that were subjected to hot water treatment. It is therefore recommended that hot water treatment of dormant nursery plants be included in an integrated strategy for the proactive management of these diseases in grapevine nurseries. 相似文献