双染的

Being a microcosm himself, he discovers — and it is a true discovery, and he is the man to make it — that the world has been eating green apples; to his eyes, in fact, the globe itself is a great green apple, which there is danger awful to think of that the children of men will nibble before it is ripe; and straightway his drastic philanthropy seeks out the Esquimaux (16) and the Patagonian,(17) and embraces the populous Indian and Chinese villages; and thus, by a few years of philanthropic activity, the powers in the meanwhile using him for their own ends, no doubt, he cures himself of his dyspepsia, the globe acquires a faint blush on one or both of its cheeks, as if it were beginning to be ripe, and life loses its crudity and is once more sweet and wholesome to live.

他是大千世界里的一个缩影，他发现，这是一个真正的发现，而且是他发现的，——世界在吃着青苹果；在他的眼中，地球本身便是一只庞大的青苹果，想起来这却很可怕，人类的孩子如果在苹果还没有成熟的时候就去噬食它，那是很危险的；可是他那狂暴的慈善事业使他径直去找了爱斯基摩人、巴塔哥尼亚人，还拥抱了人口众多的印度和中国的村落；就这样由于他几年的慈善活动，有权有势者还利用了他来达到他们的目的，无疑他治好了自己的消化不良症，地球的一颊或双颊也染上了红晕，好像它开始成熟起来了，而生命也失去了它的粗野，再一次变得又新鲜又健康，更值得生活了。

1St, Luo grain was truly the worker has made the mistake, we will correct 2 in the following sample, because in the order form, has not mentioned this model of scarf's conclusion way, therefore the factory was before pulled the way did, was sorry, we will press your request and the reference diagram manufacture pre-natal type 3, because scarf's span was 64 ", gibbed long 4 ", and in the design drawing, requested scarf's two homochromies, this was unable to achieve, therefore the factory in the permission error range, has adjusted the altitude which gibbed, guaranteed the total lengthWith matches colors correctly, please give your opinion,(the note: This section does not use translator, is only facilitates everybody understanding, the scarf is the black salt bi-color strip, also requests two is the gray, gives the size according to the order form, can only make 16 strips, therefore is impossible to achieve two homochromies, can only make 15 statures, or 17 statures only then the line) dyes the big goods gauze approximately to need for 5-7 days, afterward can prepare the pre-natal type, waited for that your reply opinion, thanks

外贸新手，第一个自己的订单，比较紧张，怕语气拿捏不好，请教各位啊：关于您的意见： 1，罗纹确实是工人做错了，我们会在后面的样品中更正 2，由于订单上，并没有提到这款围巾的收尾方式，所以工厂是按照之前打样的方式做的，抱歉，我们会重新按您的要求和参考图制作产前样 3，因为围巾的全长是64&，夹条长4&，并且设计图上，要求围巾的两头同色，这是无法做到的，所以工厂在允许的误差范围里，调整了夹条的高度，以保证总长和配色正确，请给出您的意见，(注：这段不用翻译，只是方便大家理解，围巾是黑灰双色条子，又要求两头是灰色，按照订单给出的尺寸，只能做16个条子，所以是不可能做到两头同色，只能做15个条，或17个条才行)染大货纱约需要5-7天，之后才能准备产前样，等待您的回复意见，谢谢问题补充：别整翻译工具。。悠关前途''谢谢

Transcriptional activity of ACL recombinants normalized by Renilla was significant difference with pGL3-Basic expect for construct -27 and -15 (P＜0.01). Construct -919 contains highest activity. 3 times reduction of transcriptional activity from-919 to -679bp indicated that negative regulation factors located probably in this region. The activity started on construct -73 suggested the basal promoter activity was located within the -73 to +77bp region; Transcriptional activity of IDHβrecombinants was not significantly different between the recombinant -58 and pGL3-Basic. The activity started on construct -82, decreasing with the length of the fragment up to -164 in despite of a bit of fluctuation, and kept increasing from construct -164 up to -279. Thus, the basal promoter activity was located within the -82 to +16bp region, whereas the upstream 197bp conferred maximal transcriptional activity.

采用5'端缺失策略，结合启动子预测结果，分别构建了8个ACL基因和19个IDH3β基因启动子区重组子，将其转染猪PK15细胞系，并利用荧光素酶双报告基因系统检测了它们的活性，结果表明：在所构建的猪ACL基因5侧翼序列的8个重组子中，除pGL-ACL27和pGL-ACL15外，其它重组子的荧光素酶活性与阴性对照均达到极显著水平(P＜0.01)，表明它们均具有启动子活性，pGL-ACL919活性最强，到pGL-ACL679活性下降了将近3倍，说明从-919bp到-679bp区域可能存在负的调控位点，从pGL-ACL621到pGL-ACL73活性有小幅度下降，到pGL-ACL27活性降到与阴性对照水平无显著差异，表明维持该启动子的基本活性区域位于-73bp到+77bp之间；IDH3β基因启动子活性开始于pGL-IB82，然后虽然有些波动，但是活性一直下降直到-164bp处，之后活性又开始升高，直到-279bp处活性达到最高，这些表明维持该启动子的基本活性区域位于-82bp到+16bp之间，从-82到-279之间的179bp区域具有最高的启动子活性。

Chicken IL-15(ChIL-15), Which was discovered in 2001, plays a main role in activating NK cells and CD8+ memory T cells, and may therefore have potential as a vaccine adjuvant.According to published ChIL-15 gene sequence, a pair of primers were designed and synthesized to clone ChIL-15 cDNA from ConA-activated chicken splenocytes by RT-PCR. Recombinant plasmid pUC19ChIL-15 carrying the ChIL-15 gene was constructed. The gene encoding mature ChIL-15 (mChIL-15) was amplified from pUC19ChIL-15 by PCR and cloned into pMD 18-T vector. After digested with Kpn I /Pst I , the mChIL-15 gene was subcloned into prokaryotic expression vector pPRoEX橦Ta and transformed into E.coli DH5a .The transformant was induced by IPTG, and the recombinant ChIL-15(rChIL-15) was expressed as a fusion protein with a histidine hexamer tag at the N-terminal end of the protein. Following expression, the protein was purified by ProBond?Nickel-Chelating Resin. The uninduced and induced protein lysates and the purified protein were separated by SDS-PAGE and transferred onto nitrocellulose membrane to identify rChIL-15 by Western blot. The rChIL-15 was used to immune the guinea pig to obtain rChIL-15 polyclonal antibody.

参考已发表的ChIL-15基因序列，设计合成引物，应用RT-PCR技术从刀豆球蛋白A活化的白来航鸡脾淋巴细胞中克隆ChIL-15 cDNA，将其与SmalⅠ酶切处理的pUC19载体连接，构建重组质粒pUC19ChIL-15；用PCR技术从pUC19ChIL-15中扩增出去信号肽的成熟ChIL-15(mChIL-15)基因，与pMD 18-T载体相连构建重组质粒pMDChIL-15；然后用KpnⅠ／PstⅠ双酶切下mChIL-15基因片段，定向亚克隆到经同样酶切处理的带有6个组氨酸标签的表达载体pP_RoEX~HTa中，构建重组质粒pP_RoChIL-15，对其测序确认读框正确后，转入大肠杆菌DH5α中诱导表达并进行纯化，用重组ChIL-15(rChIL-15)免疫豚鼠，制备多克隆抗体；再用HindⅢ／PstⅠ从质粒pP_RoChIL-15上切下mChIL-15基因，定向亚克隆到经同样酶切处理的杆状病毒转移载体pMelBacB中，经酶切、PCR和序列测定鉴定后，与杆状病毒线形化DNABac-N-Blue~(TM DNA共转染Sf9昆虫细胞，经三轮蚀斑纯化，构建重组杆状病毒rBac-ChIL-15，用该重组病毒感染处于对数生长期的Sf9细胞，按不同的时间收取样品，离心后对其上清和沉淀进行SDS-PAGE和Western blot分析。

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。

However, they formed themselves into line, all assisting, owing to the importance of the search; the dairyman at the upper end with Mr Clare, who had volunteered to help; then Tess, Marian, Izz Huett, and Retty; then Bill Lewell, Jonathan, and the married dairywomen - Beck Knibbs, with her woolly black hair and rolling eyes; and flaxen Frances, consumptive from the winter damps of the water-meads - who lived in their respective cottages.

但是由于事关重大，他们就都过来帮忙，一起排成一排搜查；克莱尔先生也自动过来帮忙，奶牛场老板就和他站在上边的开头；排在他们后面的是苔丝、玛丽安、伊茨·休特和莱蒂；再往后就是比尔·洛威尔、约纳森，还有已经结了婚住在各自房舍里的女工们——里面有贝克·尼布斯，她长了一头黑色的鬈发和一双滴溜溜直转的大眼睛；还有一个长着亚麻色头发的法兰西斯，她因为水草场上冬季的湿气而染上了肺病。

Cell lines were co-tranfected with bi-plasmids and cultured at 1% O〓 condition, luciferase assay demonstrated that all of the expression systems with different domains could be hypoxia-activated. GAL-CAD achieved more than 100-fold induction in A549 cell and more than 40-fold in HepG2; expression level of the system markedly increased after addition of glycin arm, almost 10-fold enhancement in HepG2 cell. Because basic expression also increased, the induction fold by hypoxia decreased.

双质粒共转染不同细胞株后，在1％O〓环境下培养24h，荧光素酶活性检测结果提示含不同结构域的表达系统均有受缺氧激活的能力，其中GAL-CAD在A549中获得100倍以上的诱导，在HepG2中获得40倍以上的诱导；加入甘氨酸臂后该表达系统的转录活性大为增强，在HepG2细胞中增强了10倍以上，但是由于基础表达也有所增加，受缺氧诱导的倍数下降。

For a big chunk of credit-card losses; the number of filings (and thus charge-off rates) would be rising again, whether

年美国个人破产法的一个改动使得破产登记急速下降，而后引起了信用卡大规模的亏损。

Eph. 4:23 And that you be renewed in the spirit of your mind

弗四23 而在你们心思的灵里得以更新