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1.
本研究旨在探明鸡恒定链(invariant chain,Ii)与内吞体转运蛋白Rab5a和Rab7b结合的结构域和在细胞内共定位的特征。首先,用PCR和基因突变技术将Ii胞浆区与跨膜区[Ii(Cyt-Tra)]、Ii CLIP (class Ⅱ-associated invariant chain peptide)-三聚体区[Ii(CLIP-TRIM)]和Ii突变体[Ii(M81-87aa)、Ii(M91-99aa)和Ii(M81-99aa)]分别插入pET-32a和pEGFP-C1构建相应的原核和真核重组质粒。其次,将构建的含有绿色荧光蛋白的重组质粒与实验室保存的含有红色荧光Rab5a和Rab7b的重组质粒共转染至人胚胎肾细胞系293 T,观察它们的共定位。将构建的原核重组质粒进行表达和纯化,最后用拉下法和免疫印迹检测Ii与Rab5a和Rab7b的结合域。结果表明,成功构建Ii结构域及Ii突变体的重组质粒。Ii(Cyt-Tra)及Ii突变体均能与Rab5a和Rab7b在细胞内共定位,而Ii(CLIP-TRIM)与空载体却不能。Ii的胞浆区和跨膜区是与Rab5a和Rab7b结合的功能结构域,而不是CLIP与三聚体区。综上所述,鸡Ii与Rab5a和Rab7b共定位和结合的区域是其胞浆区和跨膜区,而不是内质网腔区。这些结果提示Rab分子参与了Ii在胞内细胞器的转运机制,为进一步研究Ii及其载体在细胞内的转运机制和功能提供了新的途径。 相似文献
2.
Equine herpesvirus type 1 (EHV-1) is responsible for respiratory disorders, abortion and myeloencephalopathy (EHM) in horses. Two pathotypes of EHV-1 strains are circulating in the field: neurovirulent (N) and non-neurovirulent (NN). For both strains, CD172a+ monocytic cells are one of the main carrier cells of EHV-1 during primary infection, allowing the virus to invade the horse’s body. Recently, we showed that EHV-1 NN strains showed a restricted and delayed replication in CD172a+ cells. Here we characterize the in vitro replication kinetics of two EHV-1 N strains in CD172a+ cells and investigate if the replication of these strains is similarly silenced as shown for EHV-1 NN strains. We found that EHV-1 N replication was restricted to 7–8% in CD172a+ cells compared to 100% in control RK-13 cells. EHV-1 N replication was not delayed in CD172a+ cells but virus production was significant lower (103.0 TCID50/105 inoculated cells) than in RK-13 cells (108.5 TCID50/105 inoculated cells). Approximately 0.04% of CD172a+ cells produced and transmitted infectious EHV-1 to neighbour cells compared to 65% of RK-13 cells. Unlike what we observed for the NN strain, pretreatment of CD172a+ cells with histone deacetylases inhibitors (HDACi) did not influence the replication of EHV-1 N strains in these cells. Overall, these results show that the EHV-1 replication of N strains in CD172a+ cells differs from that observed for NN strains, which may contribute to their different pathogeneses in vivo. 相似文献
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4.
牛磺酸在猫营养中的应用 总被引:1,自引:0,他引:1
主要综述近年猫的牛磺酸营养,从研究方法来看衡量牛磺酸利用的方法与指标已较完善,对牛磺酸的代谢有了深入的了解,并进一步关注猫肠道菌群影响牛磺酸代谢的机理的研究;初步揭示大豆蛋白和脂肪等营养物质降低牛磺酸利用率的原因;在猫粮加工方面指出不良热处理导致的猫粮中牛磺酸利用率降低的机制,并分析了一些不适合配制猫粮的原料如米糠、大豆蛋白对牛磺酸代谢的影响;以及在猫粮中直接添加晶体牛磺酸的重要性。从文中可以看到,作为必需氨基酸的牛磺酸对于猫的生长繁殖来说十分重要。 相似文献
5.
河北坝上农田垄向对土壤风蚀的影响 总被引:3,自引:0,他引:3
在河北省坝上选择试验区 ,利用移动式风洞研究了农田起垄方向对土壤风蚀的影响。结果表明 ,农田起垄方向对土壤风蚀具有影响作用 :垄向与风向成 0°时风蚀量最大 ,90°时风蚀量最小 ;在 15m·s-1风速和 3cm表土含水率为 4 5 %的条件下 ,垄向与风向成 0°时的风蚀量为 90°时的 4 8倍。为减轻土壤风蚀危害 ,在河北坝上地区农田起垄的方向宜采用与自然风向NW (西北风向 ,315°)成 90°,即东北方向 4 5°。 相似文献
6.
石河子及周围垦区的土壤粘重且呈盐碱性,由于犁底层的存在,土质很难得到改善。而深松作业则是打破犁底层的主要农业技术措施之一。为此,论述了IZSS-600-3型振动深松鼠道犁的构造和工作原理,分析了试验因素对试验指标的影响,为机具今后在农业生产中的应用和推广提供了依据和参考。 相似文献
7.
以CP23重组蛋白为抗原建立检测微小隐孢子虫抗体间接ELISA方法的研究 总被引:3,自引:0,他引:3
摘要:以在E.coli高效表达的微小隐孢子虫子孢子表面抗原CP23为抗原,以辣根过氧化物酶(HRP)标记的山羊抗鼠IgG为二抗,建立了检测微小隐孢子虫抗体的间接ELISA方法。经检测筛选出最佳反应条件为1μg/孔纯化的E.coli表达的CP23抗原包被酶标板,用10%免血清进行封闭,以正常E.coli裂解上清液稀释待检血清。实验表明应用CP23重组蛋白作为诊断C.parvum抗原具有特异性高、抗原易纯化和成本低等特点。 相似文献
8.
M. Hühn 《Plant Breeding》1992,109(1):28-34
In many fields of application in plant breeding and crop science, ratios of two component traits X and Y are of interest (harvest index in cereals, leaf-to-stem ratio in forage legumes, height-to-diameter ratio in forest trees etc.). When selection is practised on the ratio X/Y of two traits X and Y, the experimenter may be interested in the resulting changes of both trait means. Based on improved approximations for the covariances between X and X/Y and between Y and X/Y and for the variance of X/Y the changes in the means of X and Y can be predicted by applying the regression approach from conventional selection theory. Explicit expressions for these correlated responses in X and Y when selection is practised on their ratio X/Y are derived and discussed. The different outcomes (decrease, zero change or increase) for the selection pressures on X and Y are characterized by phenotypic coefficients of variation of X and Y, phenotypic and genotypic correlations between X and Y and heritabilities of X and Y. 相似文献
9.
N. A. Khan 《Journal of Agronomy and Crop Science》1996,176(5):331-334
In a field trial conducted during 1992–93 and 1993–94, the effect of basal (B) nitrogen (N) (45 and 60 kg N ha−1 ) and foliar application (F) of water (W) or 10 kg N ha−1 and 400 or 600 ppm ethrel (E) (2-Chloro ethyl phosphonic acid) at 70 days after sowing was studied on leaf area index and dry mass at 90 days and pod number per plant, seeds per pod, 1000 seed weight, seed yield, oil content and oil vield at harvest of mustard ( Brassica juncea L. Czern & Coss.) cv. T-59. Recommended basal (B) application of 90 kg N ha (BN90 ) was used as control. On the basis of 2 year data it was found that basal application of 60 kg N and foliar spray of 10 kg N ha −1 and 600 ppm ethrel gave higher values for growth and yield characteristics and enhanced seed yield and oil yield by 12.5 and 14.8%, respectively over control BN90. 相似文献
10.
G. Tuitert Y. Hofmeester 《European journal of plant pathology / European Foundation for Plant Pathology》1994,100(1):19-53
A field experiment was set up in 1988 to study the development of rhizomania disease of sugar beet at different inoculum levels of beet necrotic yellow vein virus (BNYVV) in soil. Five, tenfold different, inoculum levels were created by addition of the approximate amounts of 0, 0.5, 5, 50 and 500 kg infested soil per ha (the latter corresponding to 0.01% v/v calculated to the tillage layer). A drip irrigation treatment was applied to study the influence of soil moisture on disease. Susceptible sugar beet, cv. Regina, was grown for three consecutive years.In the first year, root symptoms were not observed, but BNYVV-infected plants were detected by ELISA in low numbers at all inoculum levels at harvest. After late drilling in 1989, high numbers of infected plants, up to 90–100% in plots with the highest inoculum level, were detected already in June. Root symptoms were also observed from June onwards. In both these years disease incidence increased in time and was significantly influenced by the initial inoculum level. In the third year, the whole field was heavily diseased, and only for the non-irrigated plots incidence differed for different initial inoculum levels. The expression of symptoms by BNYVV-infected plants was influenced by initial inoculum level, thus by the amount and timing of primary infection.Root weight at harvest was not affected, but sugar content decreased with increasing inoculum level already in 1988, leading to a reduction in sugar yield of 10% at the highest inoculum level. In 1989, both root weight and sugar content decreased progressively with increasing inoculum level, resulting in sugar yield reductions of 11–66% (down to approximately 3000 kg ha–1) for low to high inoculum levels compared to the control. As the control plots became contaminated, all yields were low in 1990, still showing a decrease with increasing inoculum level in the non-irrigated plots, but an overall mean sugar yield of 3323 kg ha–1 for the irrigated ones.Sodium and -amino nitrogen content of the root, additional quality parameters determining extractability of sucrose, showed an increase and decrease, respectively, with increasing initial inoculum level already in the first year. The relative differences in contents compared to those from the control were largest for Na content. A significant negative correlation was found between Na (mmol kg–1 root) and sugar content (% of fresh weight); linear for 1988, exponential for 1989 and 1990.In spring 1989, the infestation of individual plots was assessed using a quantitative bioassay estimating most probable numbers (MPNs) of infective units of BNYVV per 100 g dry soil. The relationship between the MPns determined and root weight, sugar content and sugar yield at harvest could be described by Gompertz curves. The increase in disease incidence with increasing MPN in 1989 was adequately fitted with a logistic equation. 相似文献