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991.
将重组鸡痘病毒(rFPV—IFN-γ)与IBDMB43弱毒疫苗联合接种SPF鸡,研究重组鸡IFN-γ对IBD疫苗免疫效果的影响。结果显示,rFPV—IFN-γ和MB43弱毒疫苗联合免疫组(MB43+rFPV-IFN-)")及rlFN-γ和MB43弱毒疫苗联合免疫组(MB43+rIFN-γ)在免疫后第2周即可检出ELISA抗体,比MB43疫苗单独免疫组早1周。用IBDVGx株攻击后,MB43+rFPV-IFN-γ组和MB43+rIFN-γ组免疫鸡的脾只有个别淋巴细胞核浓缩、核崩解,少数滤泡萎缩变性坏死;胸腺损伤程度轻于MB43疫苗单独免疫组和非免疫对照组。免疫后1周,3个免疫组外周血CD8^+T淋巴细胞含量均显著高于非免疫对照组,CD4^+T淋巴细胞含量变化不明显;攻毒后第2周,3个疫苗免疫组CD4^+、CD8^+T淋巴细胞含量均显著升高,各组之间CD4^+、CD8^+T淋巴细胞含量无明显差异。证实,rFPV—IFN-γ和rIFN-γ可加强疫苗的细胞免疫和体液免疫应答。 相似文献
992.
构建了泛素(ubiquitin,Ub)基因与密码子优化的PRRSVGP5蛋白(optiGP5)基因融合表达质粒pIR—Ub—optiGP5。间接免疫荧光和Western—blot分析表明,重组质粒转染293T细胞后能够瞬时表达。将pIR—optiGP5和pIR—Ub—optiGP5两种质粒DNA肌肉注射免疫BALB/c小鼠后,分别检测小鼠血清中抗GP5抗体、T淋巴细胞的增殖能力以及CTL活性。结果显示,pIR—optiGP5质粒免疫组在二免后可以检测到荧光抗体,而pIR—Ub—optiGP5质粒免疫组一直未检测到抗GP5抗体,但该免疫组的T淋巴细胞增殖指数及针对GP5的特异性CTL反应数值明显高于pIR—optiGP5质粒免疫组。这表明泛素与GP5的融合表达可增强细胞免疫反应。 相似文献
993.
994.
995.
本研究经过菌落形态、染色形态、生化实验以及PCR实验,从来自湖北4个鸭场的81份病料中,分离到11株鸭疫里默氏茵。以5.0×10~8CFU/mL剂量感染健康昆明鼠和健康樱桃谷鸭,鼠和鸭分别在感染后7 h和12 h出现症状,直到感染后1周,实验动物均死亡,而对照组则未出现任何症状,表明该菌株为有毒力菌株,药物敏感实验结果显示部分细菌对某些药物已经产生耐药性。 相似文献
996.
Exergy, the thermodynamic function that represents the distance of an open system from equilibrium, is proposed as an ecological indicator for summarizing the complex dynamics occurring in a disturbed community during its recovery processes. These complex dynamics can be difficult to capture by using classical indices. In this study, we sampled macrobenthos using the BACI scheme (before versus after; control versus impact) in tidal wetlands of west Chongming Island, China, an area that has been disturbed by ecological engineering measures. Exergy storage is estimated for benthic communities. The control area is proposed as dynamic reference for estimating local exergy storage of the benthic community. Three different methods were used for estimating exergy on the basis of coefficients: (a) taken for taxa groups; (b) estimated from coding genes given for broad taxonomic groups; and (c) estimated from genome size taken as close as possible to the taxonomic level of the species, providing a basis for inferring similarities. We found a decrease of local exergy content in the disturbed area 9 days after the disturbance. Subsequently, the reference exergy of the benthic community increased (i.e. in the surrounding control area) in accordance with the proposed hypothesis regarding the dynamics of exergy storage during a system's development. Estimates of local exergy arrived at using the three methods provided comparable results. Based on this, we then discuss the feasibility of using the more readily available genome size data for estimating exergy. This result may have important implications for broader application of this indicator to biological systems. Moreover, the adjacent control samples seem to represent an appropriate dynamic reference for estimating the local exergy of disturbed communities. We found that exergy was a useful indicator that integrates the processes underlying the recovery of the benthic community after disturbance. 相似文献
997.
This study was aimed to isolate a mutant strain of porcine epidemic diarrhea virus and prepare a PEDV inactivated vaccine with high valence by suspension culture process for immunizing against PEDV effectively in China.200 small intestines and theirs contents of diarrhea piglets died of diarrhea,collected from many large-scale pig farms in China,were detected by RT-PCR and sequenced,a mutant strain of porcine epidemic diarrhea virus was selected and put on the suspension-cultured Vero cells in a 2 L reactor for virus isolation and continuous cell culture,the harvested virus suspension,which was identified and determined TCID50,was inactivated by formaldehyde and mixed with aluminum hydroxide gel adjuvant to prepare PEDV inactivated vaccine.After its physical behavior,stability viscosity,sterility test were checked out,the safety and immune efficacy were studied by immunizing the pregnant pigs and theirs piglets.The results were as follows:86 samples were detected positive in 200 samples,cytopathy occurred after the mutant strain samples screened were passaged to 5th generation,the virus suspension was harvested in 10th generation and identified as a mutant strain of PEDV,named PEDV-GF10 strain.The virus titer of harvested virus suspension was measured up to 1×108.0 TCID50/mL after concentrated.After the vaccine was checked out,the sows,40 and 25 days before delivery in experimental groups,were injected into Xuehai acupoint with 4 mL vaccine and the pigs in blank group were free of immunifications.The results showed that there were no obvious differences in the production status of the sows in experimental groups and blank group and the temperature of theirs 3-day-old healthy piglets injected different doses of vaccine,and the vaccine was safe to the sows and piglets.Forty 3-day-old piglets producted by pregnant sows in experimental groups and blank group were randomly selected and taken 4 mL 1×108.0TCID50/mL F10 virus culture.The PEDV morbidity of piglets in blank group was 100% after injection and the antibodies were negative;10% piglets in blank group had mild diarrhea symptoms,the protection rate was up to 90%,antibody of passive immunity in piglets lasted for more than 35 days.Virus titer of mutant strain of PEDV-GF10 improved a lot by suspension cell culture,the PEDV-GF10 inactivated vaccine was safe,and could effectively prevent and control the variation strain of PEDV in China. 相似文献
998.
马传染性贫血病毒(Equine infectious anemia virus,EIAV)的密码子使用频率与哺乳动物间存在着明显差异。为此,对马传染性贫血病毒驴白细胞弱毒疫苗株(DLA-EIAV)囊膜全长基因按照哺乳动物优势密码子的使用原则进行了重新设计和合成,并以此为基础通过重叠延伸PCR、限制酶切等方法得到结合型和分泌型囊膜基因,将其插入含有鸡beta-actin/兔beta-globin复合启动子(AG)的高效表达载体pCAGGS中,构建了EIAV驴白细胞弱毒疫苗株结合型和分泌型囊膜基因的DNA疫苗质粒pCAGGS-opti-bou-env、pCAGGS- opti-sec-env。将构建的质粒纯化后分别转染293T细胞,以间接免疫荧光和Western blot方法检测转染48 h后细胞及上清中囊膜蛋白的表达。结果显示,两种表达质粒均可正确表达EIAV囊膜蛋白,与相对应未优化的表达载体pCAGGS-wt-bou-env和pCAGGS-wt-sec—env相比,密码子优化的基因体外瞬时表达水平有极为显著的提高,而且蛋白表达部位也与预期的结果符合。这一结果为EIAV囊膜蛋白的单抗制备、表位鉴定、免疫试验、新疫苗的开发等奠定了基础。 相似文献
999.
马γ-干扰素成熟蛋白基因在大肠埃希氏茵中表达及其多克隆抗体制备 总被引:1,自引:0,他引:1
采用RT-PCR从ConA刺激的马外周血单核细胞(PBMC)中扩增获得含信号肽的马γ-干扰素(Equine interferon-γ,EIFN-γ)基因,将其克隆至载体pCR2.1-TOPO中。通过PCR方法从重组质粒(pCR-EIFN-γ)中扩增马干扰素成熟蛋白(MatureEIFN-γ,mEIFN-γ)基因,并将其亚克隆至原核表达载体pET-28a(+)。重组子经测序鉴定正确后转化至大肠杆菌BL21(DE3),经IPTG诱导,对其产物进行SDS-PAGE分析和Western blot鉴定,将纯化后的表达产物免疫新西兰白兔制备多克隆抗体。结果表明,mEIFN-γ基因全长为441b口,含一个开放阅读框,编码146个氨基酸的成熟蛋白;对其表达产物以可溶性和包涵体两种形式存在,重组蛋白相对分子量约为18Ku,且具有免疫反应活性。mEIFN-γ基因在大肠杆菌中高效表达并在非变性和变性条件下,采用Ni^+亲和层析纯化方法均可获得高纯度的重组马γ-干扰素制备的多克隆抗体,为建立马γ-干扰素检测方法、监测机体免疫状态和研究机体免疫机制奠定了基础。 相似文献
1000.
将猪圆环病毒2型(Type2 porcine circovirus,PCV2)JXL株cDNA插入到载体pMD18-T中,获得的pMDT-PCV转染猪肾细胞PK15细胞系,盲传4代后,利用PCV2阳性猪血清进行间接免疫荧光抗体试验(IFA),能检测到特异性荧光。将pMDT-PCV质粒DNA腹股沟淋巴结注射40日龄仔猪2头,分别接种2、3次,接种剂量500μg/次,首次注射35d心脏放血致死。在体内感染性试验期间,动物未出现明显临床症状。试验结束,病理切片结果显示,不同淋巴组织中分别存在淋巴细胞缺失或增多的现象;肾小球、扁桃体、空肠淋巴结和股前淋巴结等冰冻组织切片中存在大量PCV2病毒抗原;血清中PCV2特异性的ELISA抗体效价迭1:2560,IFA效价为1:1280;通过聚合酶链式反应(PCR)可以分别从体外感染PK15细胞和体内感染动物及同圈饲养的空白对照猪的淋巴组织中扩增到病毒特异性基因片段。本研究证明含有单拷贝PCV2的重组质粒pMDT—PCV在体内、外均具有感染性,能衍生出病毒,并产生符合自然感染PCV2的大体病变和组织病理学变化,并激发较强的体液免癌应答。 相似文献