小鼠单个冷冻胚胎银染mRNA差异显示方法的建立

为建立单个冷冻胚胎银染mRNA差异显示方法,用小鼠单个4C鲜胚和冻胚RT—PCR产物,验证了冻胚和鲜胚实验结果的一致性。并从小鼠4C和8C冻胚的银染聚丙烯酰胺凝胶中回收了8C期的特异条带,通过再扩增、同收、克隆及酶切鉴定,克隆到了小鼠8C期胚胎差异表达的小鼠RP23—20A6基因。结果表明,所研究建立的单个冻胚银染mRNA差异显示方法具有可行性和有效性,可以用于动物早期胚胎基因表达的研究。     

贮梨冷库生产金针菇时的CO2浓度变化及调控研究

针对闲置贮梨冷库进行金针菇生产易造成库内CO2过度积累而影响金针菇商品性能问题,采用观测和模拟试验,对贮梨冷库生产金针菇时的CO2浓度变化及排除方法进行了研究.结果表明:金针菇菌丝体和子实体发育过程中不断产生CO2,且所产生的CO2浓度随时间延长而增高;在冷库近底层增加新风口,与库门构成平抽气流的改建设计,能有效提高CO2的排除效果,机械通风并辅以化学吸收剂Ca(OH)2,可有效保障库内CO2浓度与金针菇子实体生长发育的不同阶段相适应.     

mRNA差异显示技术的研究进展

对mRNA差异显示技术中引物设计、应用非放射性标记、优化PCR参数、假阳性鉴定的方法等方面进行了概述。     

mRNA差异显示技术的研究进展及其在植物研究中的应用

综述了mRNA差异显示技术的原理、技术路线、优缺点以及差异显示技术的改进过程,并对其在植物发育、植物抗性机理、植物杂种优势机理以及植物激素诱导的基因表达等植物研究领域中的应用作了简要的介绍.     

梅花开花物候期及加长观赏期的研究

根据１８１个梅花品种开花物候期观察记载,在南京地区拟定２月２０日以前进入开花最佳观赏期的为早花品种;２月２０日～３月１０日进入开花最佳观赏期的为中花品种;３月１０日以后进入开花最佳观赏期的为晚花品种．早花品种最佳观赏期最长为２８ｄ,最短为１３５ｄ,平均１７４ｄ;中花品种最佳观赏期最长为１５ｄ,最短为８５ｄ,平均为１１５ｄ;晚花品种花期最长为９６ｄ,最短５ｄ,平均７８ｄ．选取５０个左右的代表品种组成梅花园景品种组合,其中早、中、晚花品种植株数量按３５∶３５∶３的比例配植,可使梅园在正常气候年景梅花最佳观赏期达到５０～５６ｄ,较目前一般梅园观赏期可加长２０～２５ｄ左右,从而大大提高梅园的观赏效益．     

细胞表面展示有机磷水解酶的恶臭假单胞菌工程菌的构建及全细胞酶活性分析

利用丁香假单胞菌(Pseudomonas syringae)冰核基因的N-末端(inpn)作为锚定单元(anchoring mo-tif),通过与来自黄杆菌属(Flavobacterium sp.)的有机磷水解酶基因(opd 构建融合基因inpn-opd,并连接于假单胞菌表达载体Pymbp,然后导入恶臭假单胞菌(P.putida)野生型菌株AB92019,获得了能在其细胞表面展示有机磷水解酶并具有全细胞酶催化活性的重组工程菌MMBL-opd.SDS-PAGE结果表明,融合基因能表达产生80 ku的蛋白质.重组菌MMBL-opd在无抗性LB固体培养基上能稳定生长,所携带的外源质粒的稳定性达到100%;在添加100 μmol/L Go2 培养基上28℃培养48 h,表面展示的有机磷水解酶具有最高全细胞酶活性,为0.036 U/mg.用蛋白酶K消化处理重组菌表面蛋白可使其全细胞酶活降低92%.重组菌在PBS缓冲液中于4℃条件下保存30 d,仍能保持93%的全细胞酶活.     

土壤重金属污染的微生物修复研究进展

从利用微生物降低重金属的毒性、微生物展示技术用于土壤重金属污染的治理、生物表面活性剂去除土壤重金属、植物根际宜生菌对重金属的修复等方面对土壤重金属污染微生物修复的研究进行了综述,以期为同类研究提供参考.     

Surface-exposed expression of Edwardsiella tarda EseB in live attenuated Vibrio anguillarum based on novel surface display systems

Live, attenuated Vibrio anguillarum strains can serve as vectors for the delivery of heterologous antigens for development of multivalent recombinant vaccines. Based on the outer membrane anchoring elements of V. anguillarum , we have previously constructed several efficient surface display systems Lpp-Omporf1, Lpp-OmpU, Lpp-Omp26La, Wza-Omporf1, Wza-OmpU and Wza-Omp26La. In this study, with these constructed surface display systems, a putative antigen protein EseB from pathogenic Edwardsiella tarda was successfully expressed on the surface of an attenuated V. anguillarum strain to get multivalent vaccine candidates. Further immune protection evaluation in zebra fish ( Danio rerio ) demonstrated that the V. anguillarum EseB-display strain AV/pW-26La-B could trigger full protection against V. anguillarum infection and early protection against E. tarda infection in the immunized fish. These results suggest that surface display of heterologous protective antigens in attenuated V. anguillarum could be used as a tool to develop potential V. anguillarum vector vaccine.     

基于DELPHI的双屏显示系统的设计与实现

计算机双屏显示是计算机技术与可视化技术相结合的一种新应用.双屏显示系统帮助提高工作效率,避免频繁切换窗口.本文提出了双屏显示的概念,并详细分析了该技术的实现细节,最后介绍了它在实际操作中的应用.     

Detection of HCV core protein,envelope glycoprotein and core-envelope fusion protein by baculovirus surface display

AIM: To detect the hepatitis C virus (HCV) at early stages by the technique of baculovirus surface display. METHODS: We constructed the recombinant baculoviruses vAc-Flag-HCV C-GP64, vAc-Flag-HCV E-GP64 and vAc-Flag-CE-GP64, which displayed HCV-related structural proteins by the technique of baculovirus surface display. RESULTS: Western blotting results showed that the core (C), envelope (E) and core-envelope (CE) proteins of HCV were expressed in Sf9 cells after the infection of recombinant baculoviruses. We also observed by confocal microscopy that the C, E and CE proteins of HCV presented and were anchored on the plasma membrane of Sf9 cells after infection. In addition, the results of immunogold electron microscopy demonstrated that the 3 recombinant baculoviruses displayed the C, E and CE proteins of HCV on the viral surface, respectively. To further define the role of these recombinant baculoviruses in detecting HCV at early stages, we applied time-resolved fluoroimmunoassay (TRFIA) to detect HCV samples after purification and concentration of the recombinant baculoviruses. The results showed that the positive rate of vAc-Flag-CE-GP64 was up to 80.2%. CONCLUSION: vAc-Flag-CE-GP64 is useful for detecting the hepatitis C virus at early stages.