上切

A branch cut off from the adjacent branch must of necessity be cut off from the whole tree also.

从邻枝上切下的一根枝条必定也是从整个树上切下的。

With primers designed according to sequence(AY052417) of diapause pupae of oriental tobacco budworm registered on GenBank, a double-strand cDNA fragment of DH gene was obtained from RT-PCR amplification and was further cloned into pGEM-T Easy vector.

根据GenBank发表序列(AY052417)设计引物，以烟夜蛾(Helicoverpa assulta Guenée)滞育蛹为材料，利用RT-PCR技术扩增到滞育激素(diapause hormone，DH)cDNA，并通过TA克隆的方法将其克隆至pGEM-T Easy载体上，对目的基因进行测序，并利用设计的酶切位点将目的基因从T载体上切下。

She put the vegetables on a chopping board and began to cut them up.

她把菜放在切菜板上切了起来。

Kabardian is also known as East Circassian or Upper Circassian.

卡巴尔达语也被称作东切尔克斯语或上切尔克斯语。

In this research we obtained the banana ACO gene with about 1.2kb length by cuffing from the cloning vector pUCm-ACO with restriction endonucleases Xba Ⅰ and Sac Ⅰ, and then directionally linked the gene to the plasmid vector pBⅠ-121 in the same endonucleases digestion sites, finally we got the construct of plant expression vector pBI-aACO.

用限制性内切酶Xba Ⅰ和sac Ⅰ从克隆载体pUCm-ACO上切下约1.2kb的香蕉ACO基因，将其定向连接在经相同酶切的质粒载体pBⅠ-121上，构建成植物表达载体pBⅠ-aACO。

The carving is done in the kitchen, and not on the dining table as in the West.

格格切肉是在厨房里进行的，而不是像西方那样在餐桌上切肉。

And then adding NAA at low concentration in MS was used to give rise to roots. We also found that lower level of hormone could control effectively browninng and vitrifaction during the culture and G1 n (6mg/L) and AgNOj (2mg/L) supplemented in shoot induction media could improve the shoot information rates apparently (about90%).The whole period of plant regeneration from leaflets of peanut could be divided as five steps: germination - shoot induction -shoot elongation-rooting- tranplant.

用1／2MS培养基萌发花生种子，9-10d后，从无菌花生苗上切取幼嫩叶片中部为外植体。2500Lux光照和27±1℃条件下，在诱芽培养基(MS+BA3mg／L+NAA0.8mg／L+AgNO_32mg／L十Gln 6mg／L)培养12-14d即可观察到明显芽点或瘤状突起，较前人报道的培养时间大大缩短了，4w后芽点进一步发育成丛生芽，芽诱导率达90.2％，每个外植体平均产9个丛尘芽，然后转至培养基MS+BA3 mg／L+AgNO_32 mg／L上诱导芽的伸长，3-4w后可长至3-4cm，切下带有2-3片叶片的幼芽移至生根培养基(MS+NAA0.8mg／L+AgNO_32mg／L)，1w后切口处可见白色不定根形成。

The results are as follows:(1) With the imposing of orthopedic force of simulating maxillary protraction on maxilla, in the sagittal direction, the maxillary complex of the SD Rats in experimental group grows more quickly than that in two control groups, and the extent of the maxillary complex increases more evidently. The maxillary complex displays anti-clockwise forward and upward rotation.(2) With the imposing of orthopedic force on maxilla, upper incisors exhibit labial-inclined, labioclination of lower incisors changes insignificantly, and the shape and the dimension of skull are not affected significantly.(3) The expression of TGF-3 , in frontomaxillary sutures and palatomaxillary sutures shows different intensity in different extent of time. But The expression of TGF-{3, in experimental group is more intensive than that in two control groups, and that in operated control group and natural growth group has no significant difference.

结果如下：在模拟上颌前牵引矫形力作用下，(1)生长发育期大鼠上颌复合体在矢状方向生长加快，长度明显增加，并表现出向前向上的逆时针旋转；(2)上切牙唇倾，下切牙的唇倾度没有明显变化，颅脑的形态与大小不受影响；(3)额颌缝和腭颌缝中TGF-β_1的表达，同一时段实验加力组要强于手术对照组与自然生长组，而手术对照组与自然生长组之间无显著性差异。

The PSCA_3 fragment was selected for its superior expression level in eukaryotic cells.Then the sig-PSCA_3-Fc-GPI genetic fragment was cloned into pVAX1-neo-IRES-GM/B7 vector to construct the final immunological inhanced DNA vaccine pVAX1-PSCA_3-FcGB. Immunofluorescence and flow cytometry were used to confirm the expression of PSCA_3 fragment by transfected into Cos7 cell.Finally,the anti-tumor effect of pVAX1-PSCA_3-FcGB was tested in murine prostate cancer model generated by RM-1 cell line.The animal was immunized with pVAX1-PSCA_3-FcGB DNA vaccine by intramuscular injection plus electroporation,pVAX1 and pVAX1-PSCA_1-FcGB plasmid were used as control.The inhibitory effect of tumor was investigated by observion of forming time,volume and inhibition ratio of tumor.Results:DNA sequencing conformed that the heterological PSCA fusion antigen fragment which was synchronized by overlapping-extending-PCR,was consistent to design.Enzyme digestion analysis showed that the 1 to 4 copies heterological PSCA fusion antigen fragments were constructed successfully.

方法(1)检索GenBank，选择包含人主要T细胞抗原表位序列的人PSCA基因片段，应用异种化抗原设计技术，保留人T细胞抗原表位，设计异种化PSCA融合抗原片段；(2)根据核酸序列按中心模板法设计引物，应用重叠延伸PCR技术拼接合成异种化PSCA融合抗原片段基因，以PCR、限制性酶切和DNA序列测定法进行鉴定：(3)利用DNA限制性内切酶BssHⅡ和MluⅠ酶切后粘端互补的特点，采用同尾酶法构建1—4拷贝异种化PSCA融合抗原片段(PNCA_1-PSCA_4)，并将上述片段分别插入真核表达载体pCI-neo-Fc-GPI中，转染293T细胞，借助免疫荧光+流式细胞术考察插入片段表达效率，最终选定PSCA_3片段进行下一步研究；(4)将sig-PSCA_3-Fc-GPI基因片段自pCI-PSCA_3-Fc-GPI质粒上切下，插入pVAX1-neo-IRES—GM/B7载体中，构建免疫增效DNA疫苗pVAX1-PSCA_3-FcGB，并应用转染Cos7细胞+免疫荧光/流式细胞术方法鉴定其在真核细胞中的表达情况；(5)给8周龄雄性C57BL/6小鼠皮下种植RM-1细胞，制备小鼠前列腺癌模型，并采用股四头肌肌肉注射+电脉冲法(Electroporation，EP)接种DNA疫苗质粒pVAX1-PSCA_3-FcGB，同时接种pVAX1空载体质粒和pVAX1-PSCA_1-FcGB质粒作为对照，通过观察计算免疫动物的成瘤时间、肿瘤体积和抑瘤率，来评价该DNA疫苗在小鼠体内的抑瘤效果。

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。

They weren't aggressive, but I yelled and threw a rock in their direction to get them off the trail and away from me, just in case.

他们没有侵略性，但我大喊，并在他们的方向扔石头让他们过的线索，远离我，以防万一。

In slot 2 in your bag put wrapping paper, quantity does not matter in this case.

在你的书包里槽2把包装纸、数量无关紧要。