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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。

Because the light power in claddings is weaker than one in core, the overlap will weaken when modulation is realized only in cladding.

由于层中光波强度比芯层中要弱,因此在层中调制光波会引起场模交叠因子的弱化,但是优化计算发现,通过调节波导的尺寸尤其是芯层厚度可以控制交叠因子使其弱化程度降低到最小,在最优的情况下,场-模交叠因子可达0.89。

This paper describes a method for embedding the sheathed thermocouples inthe aluminum claddings of some fuel elements of experimental reactors byultrasonic welding technique,and the measurement results of the claddingtemperature of fuel elements in reactors.

本文介绍一种应用超声焊接技术在几种实验堆燃料元件铝壳上埋设铠装热电偶的方法及元件壳温度随堆测量结果。

The various networks on the Internet work in pretty much the same way; they pass data around in clunks called packets, each of which carries the addresses of its sender and its receiver (those host numbers we talked about earlier).

Internet的各种网络按相同的方式工作:它们将数据块以所谓的&&来传送,每个中载有发送人和接收人的地址(即前面介绍过的主机编号)。

Lump commerces were entitled to detain or later pay subcontractor for all or part expiration crash,without damaging the right of applicable law or subcontract.

在不损害适用法律或分合同所规定的任何权利的情况下,总商有权扣留或暂缓支付总商应付给分商的全部或部分到期金额。

She imagined herself a belated traveler, a poor girl, an outcast, quenching her thirst at the wayside brook, her little packet of cresses doing duty for a bundle of clothes

她想象自己是一个日暮无归处的旅人,一个穷姑娘,一个流浪汉在路旁小溪内喝水解渴,那一小水田芹就权充衣

29The boards are to be covered with gold, and with gold rings on them to take the crossbars which you are to cover with gold.

29木板应上金,穿横木的环子应是金做的,横木也上金。

They've been mailing tea bags to Congress to I guess express their dissatisfaction with taxes and government spending because nothing shakes a politician up like a complimentary bag of tea.'Hey if you don't straighten up next year, crumpets, buddy.

他们举行了&茶党&抗议,一直在往国会寄茶,我猜他们是想表达他们对税收和政府开支的不满,因为没有什么能比一个赠送的茶更能动摇一个政客了:'嘿,伙计,要是你们明年再不改,我们就寄煎饼。

All the preparation I did minded me alot of my memory of heading for Singapore 7 years ago . Seven years! Time really can fly really fast when I realized it, Then I was also busy with packing, so many clothes I took with me and most of which are formal dress, ridiculous! Due to the fact I had worked in insurance company before I got the job in Star Crusies, almost all my clothes were suits for formal occasions.

想起7年前,也是夏天,我第一次走出国门,飞往我曾经梦想之地新加坡,也是准备大的行李,那时好傻,带了好多衣服,因为之前在保险公司工作,衣柜里全是正装,就带去了大量的套装,到了那里几乎都没穿,最后扔了一部分,剩下的那部分一年后原封不动地背回来。

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推荐网络例句

With Death guitarist Schuldiner adopting vocal duties, the band made a major impact on the scene.

随着死亡的吉他手Schuldiner接受主唱的职务,乐队在现实中树立了重要的影响。

But he could still end up breakfasting on Swiss-government issue muesli because all six are accused of nicking around 45 million pounds they should have paid to FIFA.

不过他最后仍有可能沦为瑞士政府&议事餐桌&上的一道早餐,因为这所有六个人都被指控把本应支付给国际足联的大约4500万英镑骗了个精光。

Closes the eye, the deep breathing, all no longer are the dreams as if......

关闭眼睛,深呼吸,一切不再是梦想,犹如。。。。。。