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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。

This English certificate examination system, a set of full and independent certificate system consisting of Listening, Speaking, Reading and Writing, was developed in 1999 by the University of Cambridge Local Examinations Syndicate on the basis of full market research of English Learning in over 100 non-English speaking countries. The first CELS test was administered on May 2002 throughout the world.

CELS证书考试体系，是由剑桥大学考试委员会，在对全世界一百多个非英语国家英语学习市场需求，进行充分和全面研究的基础上，为16岁以上英语为非母语者，在1999年设计建立的一套具备听、说、读、写各单项技能组成的完整和独立的英语证书考试体系，首次考试业已于2002年5月在世界各地进行。

In this free nature, why some iters were not chuffed, because of either occupational habits or mental exhaustion, or because of the cities that are full of material but also full of more and more people and that can't give people good mood.

在这轻松的自然里，为何一些 ITer 的心情似乎并没有舒展和释放，是职业的习惯，还是心理的疲惫，还是物博人更博的城市不能给人释怀。

This marks the first Pixar release that doesn't include both the widescreen and full-screen versions in the same DVD set, which was a great bargaining chip for those cinephiles who still want a full-frame presentation for other family members.

这标志著首次发布皮克斯不包括宽屏和全屏的版本在同一DVD集，这是一个伟大的筹码为那些cinephiles谁也想一全画幅介绍其他家庭成员。

We're all throwed in a world full of stils and confusions,a world full of questions and answers.

我们都处在一个充满混乱与骚动的世界，一个充满各种疑问与答案的世界。

I call this maneuver CTRL-ALT-DEL or reset. There are two buttons at the bottom of your harness, and you have to push them simultaneously to reset your glider. Whenever your glider is cravatted, and you can't get it out by picking some lines or collapsing half the wing, you use a full stall to solve the problem (but of course only if you have enough height). Sometimes a glider keeps flying great with a cravat, so if you can land it there's no need to full stall.

可以把这个动作类比为CTRL-ALT-DEL即是热启动；这里有两个&摁钮&在吊袋下面，你必须两手同时摁着你就可以将你的伞&重起&了；任何时候你进入卷夹，只要你有高度，你就可以使用全失速改出；当然，有时候卷夹并不是特别影响你的飞行，你也没有必要一定使用全失速改出，去着陆即可

The company is engaged in construction, road, municipal, culverts and bridges, construction equipment manufacturer specializing in the production of enterprises, from the business since 1993 "Chengxinruyi products, dedicated service" for the company's survival and development of the first elements, from the beginning Of a single development to the present sophisticated technical equipment, scientific rationalization of the advanced enterprise management, product research, development, design, trial production, and so on, the full realization of the computer-aided design, quality assurance product development, production and management in the full use of computer Auxiliary production management.

本公司是从事建筑、路面、市政、涵洞、桥梁、施工设备制造的专业生产企业，从1993年创业以来将&称心如意的产品、全心全意的服务&为公司生存和发展的第一要素，从起初的单一化发展到如今技术设备精良、科学合理化管理的先进企业，对产品调研、开发、设计、试制等工作，全面实现了计算机的辅助设计，保证产品的开发质量，在生产管理上全面采用微机辅助生产管理。

Another version of the full size mattress, which is the full extra-large size mattress, is only available in the Northern American region.

另一个版本的全尺寸床垫，这是充分特大尺寸床垫，仅适用于北美地区。

Main products include: XZ42 series moulding machine, TF series production line of permanent mould with resin sand facing, Z94 series full automotive core-shooting machine, Z95 series full automotive shell core making machine, ZHTO series core-shooting machine, ZJQ series tilt able pouring machine, ZHTO550A model core-shooting machine special for producing braking disc, as well as moulding plate pattern, core-shooting box, shell mould/core pattern/box, and so on.

主要产品有：XZ42系列造型线、TF系列铁型覆砂线、Z94系列全自动射芯机、Z95系列全自动壳芯机、ZHTO系列射芯机、ZJQ系列可倾式浇注机、ZHTO550A型刹车盘专用射芯机及型板模、射芯模、金属型和壳型模等等。

Do you know, i need you to come back

你知道吗，我需要你回来

Yang yinshu、Wang xiangsheng、Li decang,The first discovery of haemaphysalis conicinna.

1〕 杨银书，王祥生，李德昌。安徽省首次发现嗜群血蜱。