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A variety of techniques from molecular biology have been used in research laboratories to study the effects of DNA binding small molecules, such as the gel mobility shift assay,~1H NMR, electrochemistry analysis, DNA foot-printing assay, and fluorescence-based assays, UV-Vis spectrum and viscometry.

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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.

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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.

為研制帶有O型口蹄疫病毒(Foot-and-Mouth Disease Virus，F(xiàn)MDV)China99株結(jié)構(gòu)蛋白基因及多個(gè)非結(jié)構(gòu)蛋白基因的DNA疫曲av优选精品久久国产，本研究通過定點(diǎn)突變方法和PCR擴(kuò)增方法情欲网址，獲得包含有FMDV China99株結(jié)構(gòu)蛋白P1性生活视频一级片、非結(jié)構(gòu)蛋白2A、3C以及部分2B经典A片免费观看视频、3B編碼基因的片段P12X3C和包含有FMDV China99株結(jié)構(gòu)蛋白P1国产免费A片在线网址、非結(jié)構(gòu)蛋白2A、3C在线观看的毛片网站、3D以及部分2B日韩校园激情无码视频、3B編碼基因的片段P12X3C3D，將獲得的基因片段直接／酶切后與同樣處理的真核表達(dá)質(zhì)粒連接黑人大战白妞在线无码，分別得到重組質(zhì)粒pcDNA3.1／P12X3C和pcDNA3.1／P12X3C3D超碰人人曰人人操、pTARGET／P12X3C3D；對重組質(zhì)粒進(jìn)行序列測定久久精品无码一级二级、分析国产高清视频观看，并將重組質(zhì)粒分別轉(zhuǎn)染BHK-21細(xì)胞，通過雙抗體夾心ELISA方法和間接免疫熒光標(biāo)記方法檢測細(xì)胞中FMDV抗原的表達(dá)www.birugy.com，用電子顯微鏡觀察病毒空衣殼的組裝18+在线网站；為評價(jià)重組質(zhì)粒作為DNA疫苗對實(shí)驗(yàn)動物及本動物的免疫效果，將重組質(zhì)粒經(jīng)肌肉注射方法接種豚鼠中午字幕中文字幕在线中777，并與酵母表達(dá)的純化FMDV China99株3D蛋白及帶有牛α干擾素的真核表達(dá)質(zhì)粒pcDNA3.1／IFN分別／同時(shí)免疫宁夏暇葩货运代理有限公司，第二次免疫后第三周豚鼠攻以100ID_(50)或1000ID_(50)的O型FMDV China99株：隨后將質(zhì)粒pcDNA3.1／P12X3C、pcDNA3.1／P12X3C3D與帶有牛α干擾素的真核表達(dá)質(zhì)粒pcDNA3.1／IFN同時(shí)免疫牛无码一区，三周后經(jīng)牛舌皮攻以10~4ID_(50)的O型FMDV China99株秋葵视频安卓手机下载。