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By use of site mutation strategy and PCR technology, we obtained the gene P12X3C that includes full length P1, 2A, 3C and a part of 2B and 3B and the gene P12X3C3D that includes full length P1, 2A, 3C, 3D and a part of 2B and 3B. After being digested by restriction enzyme respectively, the gene P12X3C and the gene P12X3C3D were cloned into the pcDNA3. 1 and pTARGET expression vector that were digested by the same enzyme. Recombinant plasmids were checked by restriction enzyme analysis and nucleic acid sequencing. Further more, recombinant plasmids were transfected into BHK-21 cells by using lipoid. The proteins of foot-and-mouth disease virus , which were expressed in BHK-21 cells, were confirmed by sandwich-ELISA and fluoroscopy, and the capsid of FMDV was tested by electron microscope. In order to evaluate enhanced immune response of guinea pigs against FMDV, DNA vaccines which were designed to produce viral capsids lacking infectious viral nucleic acid and contained the gene P12X3C and the gene P12X3C3D were injected respectively with FMDV 3D protein which was expressed in Pichia Pastoris Secreted expression System and purified or with pcDNA3. 1/IFN which includes the gene IFN-α of cattle. Subsequently, Recombinant plasmids were injected to cattles with or without pcDNA3. 1/IFN. Anti-FMDV antibodies were detected by ELISA, and the T lymphocyte proliferation response was tested by MTT assay, neutralization antibodies titers were analyzed by micro-neutralization assay.

为研制带有O型口蹄疫病毒(Foot-and-Mouth Disease Virus，FMDV)China99株结构蛋白基因及多个非结构蛋白基因的DNA疫苗，本研究通过定点突变方法和PCR扩增方法，获得包含有FMDV China99株结构蛋白P1、非结构蛋白2A、3C以及部分2B、3B编码基因的片段P12X3C和包含有FMDV China99株结构蛋白P1、非结构蛋白2A、3C、3D以及部分2B、3B编码基因的片段P12X3C3D，将获得的基因片段直接/酶切后与同样处理的真核表达质粒连接，分别得到重组质粒pcDNA3.1/P12X3C和pcDNA3.1/P12X3C3D、pTARGET/P12X3C3D；对重组质粒进行序列测定、分析，并将重组质粒分别转染BHK-21细胞，通过双抗体夹心ELISA方法和间接免疫荧光标记方法检测细胞中FMDV抗原的表达，用电子显微镜观察病毒空衣壳的组装；为评价重组质粒作为DNA疫苗对实验动物及本动物的免疫效果，将重组质粒经肌肉注射方法接种豚鼠，并与酵母表达的纯化FMDV China99株3D蛋白及带有牛α干扰素的真核表达质粒pcDNA3.1/IFN分别/同时免疫，第二次免疫后第三周豚鼠攻以1OOID〓或1000ID〓的O型FMDV China99株；随后将质粒pcDNA3.1/P12X3C、pcDNA3.1/P12X3C3D与带有牛α干扰素的真核表达质粒pcDNA3.1/IFN同时免疫牛，三周后经牛舌皮攻以10〓ID〓的O型FMDV China99株。

By use of site mutation strategy and PCR technology, we obtained the gene P12X3C that includes full length PI, 2A, 3C and a part of 2B and 3B and the gene P12X3C3D that includes full length PI, 2A, 3C, 3D and a part of 2B and 3B. After being digested by restriction enzyme respectively, the gene P12X3C and the gene P12X3C3D were cloned into the pcDNA3.1 and pTARGET expression vector that were digested by the same enzyme. Recombinant plasmids were checked by restriction enzyme analysis and nucleic acid sequencing. Further more, recombinant plasmids were transfected into BHK-21 cells by using lipoid. The proteins of foot-and-mouth disease virus, which were expressed in BHK-21 cells, were confirmed by sandwich-ELlSA and fluoroscopy, and the capsid of FMDV was tested by electron microscope. In order to evaluate enhanced immune response of guinea pigs against FMDV, DNA vaccines which were designed to produce viral capsids lacking infectious viral nucleic acid and contained the gene P12X3C and the gene P 12X3C3D were injected respectively with FMDV 3D protein which was expressed in Pichia Pastoris Secreted expression System and purified or with pcDNA3.1/lFN which includes the gene IFN-a of cattle. Subsequently, Recombinant plasmids were injected to catties with or without pcDNA3.1/IFN. Anti-FMDV antibodies were detected by ELISA, and the T lymphocyte proliferation response was tested by MTT assay, neutralization antibodies liters were analyzed by micro-neutralization assay.

为研制带有O型口蹄疫病毒(Foot-and-Mouth Disease Virus，FMDV)China99株结构蛋白基因及多个非结构蛋白基因的DNA疫曲，本研究通过定点突变方法和PCR扩增方法，获得包含有FMDV China99株结构蛋白P1、非结构蛋白2A、3C以及部分2B、3B编码基因的片段P12X3C和包含有FMDV China99株结构蛋白P1、非结构蛋白2A、3C、3D以及部分2B、3B编码基因的片段P12X3C3D，将获得的基因片段直接／酶切后与同样处理的真核表达质粒连接，分别得到重组质粒pcDNA3.1／P12X3C和pcDNA3.1／P12X3C3D、pTARGET／P12X3C3D；对重组质粒进行序列测定、分析，并将重组质粒分别转染BHK-21细胞，通过双抗体夹心ELISA方法和间接免疫荧光标记方法检测细胞中FMDV抗原的表达，用电子显微镜观察病毒空衣壳的组装；为评价重组质粒作为DNA疫苗对实验动物及本动物的免疫效果，将重组质粒经肌肉注射方法接种豚鼠，并与酵母表达的纯化FMDV China99株3D蛋白及带有牛α干扰素的真核表达质粒pcDNA3.1／IFN分别／同时免疫，第二次免疫后第三周豚鼠攻以100ID_(50)或1000ID_(50)的O型FMDV China99株：随后将质粒pcDNA3.1／P12X3C、pcDNA3.1／P12X3C3D与带有牛α干扰素的真核表达质粒pcDNA3.1／IFN同时免疫牛，三周后经牛舌皮攻以10~4ID_(50)的O型FMDV China99株。

In South Africa's Minister of Health and others versus Treatment Action Campaign and others, 2002, the Treatment Action Campaign, having teamed up with other NGOs such as legal resource center, children's rights center, the Community Law Centre, the Institute for Democracy in South Africa and Cotlands baby sanctuary, protected the rights and interests of AIDS patients and susceptible groups by transporting low-cost AIDS drugs, oppose pharmaceutical association's attempt to stop legislation supporting affordable AIDS drugs and demanding the government to make the drug nevirapine available at all medical facilities to prevent mother-to-child HIV transmission.

南非2002年卫生部长等诉治疗行动运动组织等（Minister of Health and Others v Treatment Action Campaign and Others）案件中，治疗行动运动组织联合其他非政府组织如：法律资源中心、儿童权利中心、社区法律中心、南非民主学会以及Cotlands婴儿避难所等，通过运送低价艾滋病药物、反对医药制造协会通过诉讼的手段阻止立法对于普通的和较廉价的艾滋病药物的支持，要求政府在所有国家医疗机构中提供Nevaripine这种被广泛推荐用来减少母婴之间传播的抗逆转录病毒药物等方式，来维护艾滋病患者和易受艾滋病感染群体的利益。

Smilax, there are the phylogenetic relationships and distributive pattern to be found based on the combined ITS and maiK. tree: 1 a clade with the African species (S. kraussiana) and Australian species (S. australis), being a sister group; 2 a clade with the S. zeylanica of sect. Macranthae and S. bona-nox of sect. China, the former from Southeast Asia and latter native to US, showing they are a sister species which is disjunctive between Asia-North America, similar to the species pair of S. hispida and S. sieboldii; 3 S. perfoliata from Indonesia being sister to S. ocreata of China and S. megacarpa of Indonesia, which are very similar to each other in morphology; 4 the three species from China (S. lanceifolia, S. perfoliata and S. chapaensis) and S.ovalifolia from Indonesia showing a 87-92% bts monophyletic clade; 5 S.bracteata from China and two species of Indonesia (S. leucophylla and S. sp.) forming a clade in the ITS tree; 6 S.

通过比较研究圆锥花序组和穗菝葜组的地理分布，基于ITS及matK联合数据构建的系统树发现下列明显的具系统发育关系的类群及地理分布模式：1来自非洲的S.kraussiana和来自澳大利亚的S.australis是一对姐妹群，具有共同的祖先；2圆锥花序组的S.zeylanica和菝葜组的S.bona-nox形成一个紧密的分支，前者分布于东亚而后者只局限于美国，显示了一种类似于S.sieboldii和S.hispida那样的东亚—北美间断分布模式；3印度尼西亚的S.perfoliata是分布于中国的S.ocreata和来自印尼S.megacarpa的姐妹群，这三者在形态上也非常接近，表明具有共同祖先；4来自中国的三个种(S.lanceifolia，S.perfoliata and S.chapanensis)和来自印尼的S.ovalifolia形成了一个高支持率的单系分支(87-92％bts)；5基于ITS的系统树，来自中国的S.bracteata和来自印度尼西亚的两个种S.leucophylla and S.sp。

Main objectives of job : Selling services and products to libraries in China market Main duties and responsibilities : Understand fully the services and products that the Company can provide to library customers Actively build connections with potential customers and understand their needs, and promote and sell products or services accordingly Maintain and develop active sales channels with subagents Keep well-informed of the market trends and feedback timely to the Management Work closely with internal team in streamlining customer service and developing new product/service Requirement At least 2 years sales experience Good communications skills Reasonable English level Self-motivated hard worker Team player Location: Beijing

Frequent travel within China and infrequent travel internationally 工作任务：向中国地区的图书馆销售产品及服务主要职责：充分了解公司能为图书馆客户提供的产品和服务积极与潜在客户建立联系，了解客户需求，并按照客户的需求向其推销适合的产品或服务积极开发并维护与代理合作的销售渠道密切关注市场发展趋势，并及时向上级反馈和提供建议紧密配合内部团队完善服务质量以及开发新产品和服务职位要求：两年以上销售工作经验良好的沟通技巧良好的英语基础工作积极、主动、刻苦，富有进取心具备团队精神工作地点：北京或上海

Main objectives of job : Selling services and products to libraries in China marketMain duties and responsibilities : Understand fully the services and products that the Company can provide to library customers Actively build connections with potential customers and understand their needs, and promote and sell products or services accordinglyMaintain and develop active sales channels with subagents Keep well-informed of the market trends and feedback timely to the ManagementWork closely with internal team in streamlining customer service and developing new product/serviceRequirement At least 2 years sales experienceGood communications skillsReasonable English level Self-motivated hard workerTeam playerLocation: Beijing

Frequent travel within China and infrequent travel internationally工作任务：向中国地区的图书馆销售产品及服务主要职责：充分了解公司能为图书馆客户提供的产品和服务积极与潜在客户建立联系，了解客户需求，并按照客户的需求向其推销适合的产品或服务积极开发并维护与代理合作的销售渠道密切关注市场发展趋势，并及时向上级反馈和提供建议紧密配合内部团队完善服务质量以及开发新产品和服务职位要求：两年以上销售工作经验良好的沟通技巧良好的英语基础工作积极、主动、刻苦，富有进取心具备团队精神工作地点：北京或上海

However, the estimation results show that most estimated values are not statistically significant. According to the Burgstahler and Dichev (1997), Mills and Newberry (2001), and Phillips, Pincus, and Rego (2003) studies, this study tries to examine whether the income tax expense/benefit has the ability to forecast the earnings management by using the temporary differences between GAAP and tax law while under the control of non-discretionary accruals and tax incomes.

延伸Burgstahler and Dichev(1997)、Mills and Newberry(2001)及Phillips，Pincus，and Rego(2003)的研究，本研究试著由GAAP与税法规定同所产生之暂时性差入，在影响课税所得的情况，并控制非裁性应计项目后，检定递延所得税费用/是否对盈余管有预测能。

Nested PCR analysis and southern hybridization analysis showed that no polymorphism between H.villosa, Pm97034 and susceptible parent Wan7 107 was detected with the clones from 6VS arm, whereas three clones from H.villosa genome DNA: RH42, RH55, and RH66 showed polymorphism. RGA6 cloned from H.villosa genome DNA was characterized identity to NBS, and show high homology to resistance genes as RPM1 and RPP13 in Arabiadopsis, LRR19 in wheat, and I2C-1 in tomato.cDNA library constructed from T.aestivum-H.villosa translocation line Pm97034 was screened by hybridization with RGA6. Four positive cDNA clones were obtained. R3-2-2 showing 60-90% identity with eukaryotic sulfite oxidases contained an open reading frame of 647bp encoding 140 amino acid, and contained a conserved Moco-dimer domain in the ORF. R6-2-2 showed an ORF containing an Euk-porin domain of 279aa with 20-40% identity to the eukaryotic voltage-dependent anion channel proteins. R8-1-1 showed a complete ORF of 1810bp encoding 493aa with 80-90% identity to plant catalases. R9-1-1 showed an ORF containing a BTB/POZ conserved domain of 204aa with 30-70% identity to pox virus and zinc finger proteins. R3-2-2 and R9-1-1 were the first cDNA clones containingconserved domain of SO and POZ respectively isolated from wheat. R8-1-1 contained a complete ORF with 81% identity to Cat-3. Aspects of the role of R8-1-1 may be same with Cat-3, and it would offer the opportunity for improvement of stress tolerance of wheat.

以RGA6为探针，筛选用抗白粉病小麦—簇毛麦易位系Pm97034构建的cDNA文库，得到了4个阳性克隆。R3-2-2与动植物的亚硫酸氧化酶(sulfite oxidase，SO)有60～90％的同源性，长度为647bp，编码140个氨基酸，具有开放阅读框并含有保守域Moco-dimer.R6-2-2与真核生物的VDAC(voltage-dependent anion chennel)蛋白有20～40％的同源性，长度为1047bp，编码279aa，是一个不完整的开放阅读框，预测结构具有Euk-porin结构域。R8-1-1与植物中已经克隆的过氧化氢酶有80～90％的同源性，长度为1810bp，编码493aa，具有完整的开放阅读框和过氧化氢氧化酶保守域。R9-1-1与动植物的POZ(pox virus and zinc finger protein)蛋白有30～70％的同源性，长度为1446bp，具有开放阅读框和BTB／POZ保守域。R3-2-2与R9-1-1是首次从小麦中克隆到的具有SO和POZ保守域的cDNA序列。R8-1-1具有完整的开放阅读框，与玉米Cat-3基因具有81％同源性，预测R8-1-1可能具有与Cat-3类似的功能，可为转基因小麦抗逆育种提供新的基因。

Study engineering drawings and blueprints, determine the materials required, and plan the sequence of tasks to cut metal most efficiently · Construct patterns and templates as guides for layouts · Lay out reference points and patterns on heavy metal according to component specifications · Rig, hoist and move materials to storage areas or within worksite · Assemble and fit metal sections and plates to form complete units or subunits using tack welding, bolting, riveting or other methods · Set up and operate various heavy-duty metal-working machines such as brake presses, shears, cutting torches, grinders and drills, including computer numerical control equipment, to bend, cut, form, punch, drill or otherwise form heavy-metal components · Install fabricated components in final product.

研究机械制图和图纸，决定所需材料，并计划工作顺序，有效地切割金属。 2。构建模式和模板作为布局向导 3。按照组件规格在重金属上设计参考点和模型 4。装备、起重机和运输材料入库或在工地场内 5。组装和安装金属部件和板材形成全套装置，或通过点焊、螺栓连接，铆接法或其他方法形成次单元。 6。建立并操作各种重金属加工机床，例如压弯机、剪床、割炬、磨钻机，包括数控设备，压弯、切割、成型、打洞、钻孔，或相反形成重金属组件。 7。在成品上安装已制造好的组件 Structural metal and platework fabricators and fitters 结构金属和板金加工制造工和安装工

Main physiological and biochemical parameters were studied in different mature stage and during ripening and senescence of tomato and strawberry fruits at pink stage after harvest under low and normal temperature.

中文题名Ca2+、CaM及其目标酶与乙烯诱导番茄和草莓果实成熟衰老关系的研究副题名外文题名 Studies of relationship between Ca2+，CaM，its target enzymes and ethylene inducing maturation，ripening and senescence of tomato and strawberry fruits 论文作者顾采琴导师席玙芳李广敏学科专业食品科学研究领域\研究方向学位级别博士学位授予单位浙江大学学位授予日期2003 论文页码总数117页关键词果实植物生理学番茄草莓乙烯馆藏号BSLW /2003 /Q945 /34 研究了跃变型果实番茄和非跃型果实草莓不同成熟度以及粉红期两种果实采后置于低温和常温下后熟衰老过程中与成熟衰老有关的主要生理生化指标的变化。

In the negative and interrogative forms, of course, this is identical to the non-emphatic forms.

。但是，在否定句或疑问句里，这种带有"do"的方法表达的效果却没有什么强调的意思。

Go down on one's knees;kneel down

屈膝跪下。。。下跪祈祷