牛

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。

The fusion DNA fragments of ag85b-mpb64 and ag85b-mpb64-esat-6 were obtained by PCR andSOE technique. Various DNA vaccines were constructed with the pcDNA3.1: fusion of two genes, and of three genes, bivalent combinations and trivalent combinations(pCA+pCM+pCE6). BALB/c mice were vaccinated with this DNA vaccines.The mice injected withBCG were positive control and the mice injected with pCDNA3.1 and PBS were negative control.The mice were immunized 3 times with 2-wk intervals. The animals in group BCG were only inoculatedsubcutaneously with 1×10~6 CFU BCG at initial vaccination. The serum IgG titers and IgG isotype weredetermined using iELISA coated with M. bovis PPD and rMAE protein expressed and depurated inprokaryotic expression system every week.

同样，利用PCR和SOE技术，获得牛分枝杆菌mpb64-ag85b和mpb64-ag85b-esat-6融合基因，以pCDNA3.1为载体构建了牛分枝杆菌多价组合和多基因融合DNA疫苗：二基因融合(pCDNA3.1-MPB64-Ag85B，简称pCMA)和三基因融合(pCDNA3.1-MPB64-Ag85B-ESAT-6，简称pCMAE)DNA疫苗；二价组合和三价组合(pCA+pCM+pCE6)DNA疫苗，免疫BALB／c小鼠，以牛分枝杆菌BCG免疫组为阳性对照，以pCDNA3.1及PBS免疫组为阴性对照，共免疫3次，每次间隔2周，BCG组仅初免时皮下免疫1次。1免后每周，以原核表达纯化的重组MPB64-Ag85B-ESAT-6蛋白和牛分枝杆菌PPD为包被抗原，以间接ELISA方法检测血清IgG水平及lgG亚类。

Yeah, who doesn't like fighting cows running around with halberds?

Yeah~谁不爱牛牛呢~特别是拿着方天画戟围着你转，moo moo叫的牛牛们

In this study, we established the specialized raising model of reserved heifers, which can make weaning and mating earlier, decreasing the cost of feeding and management, increasing the quality of the whole cattle and the technique level of production management, and raising the returns of the specialized raising farm of reserved heifers.

本研究建立了后备牛专业化饲养的养殖模式，该模式的研究实施，使受试牛群犊牛断奶时间提前、育成牛配种时间提前，降低了转群成本和饲养管理费用，提高了奶牛场牛群整体水平与生产管理技术水平，并可使后备牛专业化饲养场的效益明显增加。

Tonight, by about 8:00 p.m., the waning gibbous moon and the constellation Taurus the Bull will appear together over your eastern horizon. Thereafter, the moon will shine in front of the Bull for the rest of the night.The radiant points for two November meteor showers – the South Taurids and North Taurids – both reside in front o...

我想不是说两个一模一样的星座，流星雨的名称是根据其辐射点所出现的星座范围而定的，出现在金牛座南边的就叫南金牛座流星雨，出现在金牛座北边的就叫北金牛座流星雨，出现在猎户座范围内的就叫猎户座流星雨。。。

He has finished 3 piano concerto by Mozart, 12 etudes and 10 nocturnes by Chopinin and representative pieces by Beethoven, Mendelssohn and Liszt in recent 3 years.

他4岁学琴，有着一双能够弹9度的大手，双耳可以同时听出10个音。2003年8月，牛牛在厦门举办了个人独奏音乐会，立即引起轰动，一千多位听众分享了他美妙的音乐。3年来，在两位老师的谆谆教诲下，牛牛进步神速，已弹完了3首莫扎特协奏曲，12首肖邦练习曲，10首肖邦夜曲，巴赫戈尔登堡变奏曲、以及贝多芬、孟德尔松、李斯特、斯卡拉蒂等其他音乐家的大量重要作品。

Together with the cited sequences of other species (cattle, zebu cattle and buffalo), the molecular phylogenetic tree was therefore built to probe phylogenetic relationship between Bovine subfamilies, Total 5 variation sites were examined and two types of mutation, transition and transversion, were observed, with the overage percentage of nucleotide variation of 1.66%. It demonstrated the poverty of polymorphism at exon 5 of GH gene of three bovine species. There was only one missense mutation out of 5 mutation sites which led to the shift between leu and val amino.

结果表明，在雷琼牛、巴州牦牛和巴州蒙古牛中共检测到5个变异位点，核苷酸取代方式仅有转换和颠换，核苷酸平均突变率仅为1.66%，说明这3个牛种GH基因外显子5多态性较为贫乏；5个变异位点中仅有1个为错义突变，导致亮氨酸和缬氨酸的转换，但该突变在牛种中分布有差异。

Forth, cows were immunized infected bovine rotavirus vaccine, and then the effect of passive antibody from colostrums on immunity to bovine rotavirus infections in neonatal calves was investigated.

最后，将研制的牛轮状病毒灭活疫苗免疫孕牛后，研究产下犊牛通过母源抗体获得的被动免疫在预防牛轮状病毒感染中的作用。

Animal husbandry is the ordinary people in Shandong Jiaxiang county by county Animal Husbandry Bureau approval to the improvement of cattle and sheep breeding, and optimize the culture, science and technology promotion, improved allocation in large-scale integrated animal husbandry science and technology companies, the main varieties are: Luxi cattle, Simon Tartu cattle, Limousin cattle, Charolais cattle; Boer goat, Small Tail Han sheep, Dorper sheep, the Asian Huangyangchuan, Charolais sheep.

山东老百姓牧业公司是由嘉祥县政府、县畜牧局批准，以牛羊的改良繁育、优化养殖、科技推广、良种调拨于一体的大型科技牧业公司，主要的品种有：鲁西黄牛、西门塔尔牛、利木赞牛、夏洛莱牛；波尔山羊、小尾寒羊、杜泊羊、亚洲黄羊、夏洛莱羊。

Neither the killing of Mr Zarqawi nor any breakthrough on the political front will stop the insurgency and the fratricidal murders in their tracks.

在对危险的南部地区访问时，他斥责什叶派民兵领导人对中央集权的挑衅行为。

In fact,I've got him on the satellite mobile right now.

实际上 我们已接通卫星可视电话了