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病毒脂肪酸的定性和定量,可作为病毒分类鉴定的辅助证据。有关细胞脂肪酸的研究已有报道[1~2],但对病毒多角体(polyhedrons)和颗粒体(granules)中脂肪酸成份的研究尚未见报道。本研究在病毒单糖分析的基础上[3],用毛细管气相色谱法,对... 相似文献
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杆状病毒部分功能基因的研究进展 总被引:3,自引:0,他引:3
杆状病毒以核型多角体病毒NPV的研究最为深入,到目前已有三种NPV的基因组全部测序:苜蓿银纹夜蛾核型多角体病毒(AcMNPV)[1]、黄杉古毒蛾核型多角体病毒(OpMNPV)[2]、家蚕核型多角体病毒(BmNPV)(GenBank accession no.L33180)。 杆状病毒基因组除编码病毒复制必需的基因及结构基因外,还编码一些有利于病毒充分增殖但对病毒复制并非一定必需的基因,这些基因在病毒的增殖过程中执行特定的功能,这类功能基因体现了杆状病毒的复制策略、病毒与细胞的相互作用关系,与杆状病毒宿主特异性的决定有关。本文综述了杆状病毒部分功能基因的研究进展。 相似文献
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IL—2重组杆状病毒载体的构建及表达 总被引:1,自引:0,他引:1
将人白细胞介素2(IL-2)cDNA插入苜蓿银纹夜蛾核型多角体病毒(AcN-PV)的转移载体pVL1392中。经限制性酶切和DNA杂交筛选、鉴定出重组转移载体pVL1392-IL2。重组载体通过在昆虫细胞内共转染将IL-2cDNA转移到野生型AcNPVDNA中。经32P标记探针鉴定出重组病毒Ac1392-IL2。用重组病毒感染昆虫细胞,结果表达产物有明显刺激CTLL细胞生长,[3H]-TdR掺入法检测IL-2活性为1500IU/m1。 相似文献
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核型多角体病毒属杆状病毒科,主要感染鳞翅目、双翅目和膜翅目的昆虫,具有90~160kb长的双链,超螺旋DNA基因组。现已发现约500种核型多角体病毒,这些病毒之间有什么关系?它们的亲缘关系如何?我们以甘兰夜蛾核型多角体病毒(简称MbNPV)、甜菜夜蛾核型多角体病毒(简称SeNPV)和斜纹夜蛾核型多角体病毒(简称SINPV)为材料,用限制性内切酪和分子杂交技术对它们的基因组DNA进行比较研究,并对其中一种病毒MbMV多角体蛋白基因进行了定位,现将结果报道如下;材料和方法1材料甜菜夜蛾和斜纹夜蛾健康幼虫,病毒麦种MbNPV、SeN… 相似文献
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构建家蚕Bombyx mori肌动蛋白(BmA3)启动子驱动的家蚕核型多角体病毒(BmNPV)多角体基因(ph)和OpNPV极早期启动子(IE1)驱动的zeocin抗性筛选基因转座供体载体,与鳞翅目辅助转座质粒pie2piggyBac共转染家蚕卵巢细胞BmN,经200μg/ml zeocin抗生素筛选一个月,成功获得持续表达BmNPV多角体蛋白的稳定细胞系BmN-A3ph。多角体缺陷型重组病毒BmBac-GF P感染拯救细胞系BmN- A3ph, 细胞成功装配出病毒包涵体颗粒,其包装效率约为野生型病毒感染正常BmN细胞的8%。用拯救型包涵体病毒颗粒喂食家蚕幼虫进行复感染,结果表明稳定细胞系所包装的包涵体病毒与野生型病毒一样能够通过口服途径感染宿主,却并不在宿主体内形成包涵体,从而保证外源基因高效表达。拯救型包涵体病毒可望解决传统注射感染效率较低问题,通过喂食感染可促进杆状病毒介导的家蚕生物反应器产业化进程。 相似文献
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美国白蛾核型多角体病毒实时荧光定量PCR检测方法的建立及应用 总被引:1,自引:0,他引:1
为了建立对昆虫核型多角体病毒(nucleopolyhedrovirus)进行定量检测方法,本研究选用美国白蛾Hyphantria cunea核型多角体病毒的polyhedrin基因为目的基因,经引物设计、PCR 扩增、目的片段与载体连接转化以及重组质粒的鉴定,并对重组质粒标准品和样品DNA进行检测,构建出美国白蛾核型多角体病毒SYBR GreenⅠ荧光定量标准曲线。统计学分析显示标准品浓度的对数值与Ct 值之间存在良好的线性关系(R=0.998)。该方法的检测灵敏度为101~102拷贝/μL,线性范围达5个数量级,扩增产物形成单一的特异性熔解峰,组内组间的变异系数均小于3%。根据已建立的方法对不同感染时间段的感病幼虫进行检测,每毫克幼虫样本内病毒polyhedrin基因拷贝数增殖倍数对数值与感染时间(d)呈正相关(R=0.987)。这些结果表明,建立的美国白蛾核型多角体病毒荧光定量 PCR检测方法是可靠的,具有灵敏度高、重复性好等特点,可为昆虫核型多角体病毒体内增殖动态研究及昆虫病毒的标准化生产提供参考。 相似文献
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K. Hausmann 《Protoplasma》1979,100(2):199-213
Summary The membranes of the pellicle of the ciliatePseudomicrothorax dubius are investigated using thin section electron microscopy and freeze-fracture replicas. The plasma membrane is covered by a surface coat and is connected to the outer alveolar membrane by short, sometimes branched, bridges. The inner alveolar membrane is coated on both sides. The epiplasm lies in intimate contact with the cytoplasmic surface of this membrane, and there is a corresponding deposit on the other surface. This deposit is regularly striated.The epiplasmic layer and the alveoli are interrupted at sites of cytotic activity,e.g., the attachment sites of trichocysts, the cytoproct, and the parasomal sacs. The striated deposit ends where the epiplasm ends, indicating a direct relationship between these two epimembranous layers.There is a deposit along the sides of the first part of the tip of the trichocysts, and in this region the trichocyst membrane is free of intramembranous particles.The membrane of the parasomal sacs has a coat on both surfaces. That on the extraplasmic surface is similar to the surface coat of the plasma membrane. The origin of the cytoplasmic coat is unknown. The cytotic activity of these sacs is indicated by their highly irregular profiles. 相似文献
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Summary The differentiation of the spermatid, especially in reference to the formation of the flagellum, and transformation of the shape of the nucleus was investigated in the domestic fowl.In the early stage of the spermatid, a prominent Golgi apparatus appears around the centrioles. The Golgi vesicles then surround the axial-filament complex which develops from the distal centriole. These vesicles fuse to form continuous membrane at the earliest stage of flagellar formation, and in the succeeding stage Golgi lamellae are attached to the plasma membrane of the developing flagellum. From these observations, it is assumed that Golgi apparatus may be a source of the membrane system of the flagellum.The microtubules distributed around the nucleus form the circular manchette. The anterior region of the nucleus with the manchette is cylindrical in shape and the posterior region without it remains irregular in shape. When the circular manchette has been completed, the whole nucleus acquires a slender cylindrical shape. The circular manchette then changes into the longitudinal manchette. The nuclei of spermatids without a longitudinal manchette are abnormal in shape. In view of these observations it is assumed that the nuclear shaping of the spermatid may be accomplished by circular manchette and the maintenance of shape of the elongated nucleus by longitudinal manchette.The authors wish to thank Mr. Takayuki Mori for his helpful suggestions and technical advices 相似文献
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Panova IG Sharova NP Dmitrieva SB Levin PP Tatikolov AS 《Comparative biochemistry and physiology. Part A, Molecular & integrative physiology》2008,151(4):676-681
This study aimed to analyze the aqueous humor (AH) and the vitreous body (VB) of the eye of the adult frog Rana temporaria L. as a representative species of amphibians, which lead a semi-terrestrial life. The presence of collagen, albumin, uric acid and electron donors was shown in both media; however, there are slight differences in their concentrations. To determine collagen, a spectral-fluorescent probe, cyanine dye, was used. The presence of collagen in AH of the frog was found at the first time. The total content of electron donors (ascorbic and uric acids, tryptophan, and tyrosine) in VB and HA was roughly estimated at ~ 1.5 × 10− 4 mol/L. Both VB and AH absorb light in similar UV regions. The total protein and albumin contents in AH were found to be somewhat higher than those in VB. The uric acid content was at an equally low level in both intraocular media. It is supposed that the similarity of VB and AH compositions shown in this work is due to some exchange between VB and AH contents in the course of accommodation. The role of intraocular fluids in physiological functions of the eye and in protecting the retina against UV light is discussed. 相似文献
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Summary The choriocapillaris is a fenestrated capillary bed located posterior to the retinal pigment epithelium. It serves as the main source of supply to the photoreceptors, retinal pigment epithelium, and other cells of the outer retina. The permeability of these capillaries to intravenously injected ferritin (MW — approx. 480,000; mol. diam. 11 nm) was examined in the mouse, rabbit, and guinea pig, each of which is characterized by a different type of retinal vascularization. In all three species, the bulk of the ferritin remained in the capillary lumina, where it appeared to be blocked at the level of the diaphragmed fenestrae. Some ferritin was present in endothelial cell vacuoles. The results confirm previous work on the rat choriocapillaris and indicate that the barrier function of the choriocapillary endothelium is present even among species in which the retinal circulation differs significantly.Supported by NIH grant EY03418 相似文献