克隆

As we said, Dolly was born and has been cloned by using the cell from a part of the body. It is much harder to use the cell from the body than the ovum.

就像我们所说的那样，多利的出生和克隆用的是身体某一部分的细胞，这比采用卵细胞困难得多。

The first step of the plant pterin branch is catalyzed by GTP cyclohydrolase Ⅰ and the first step in the conversion of chorismate to pABA is mediated in plants by aminodeoxychorismate synthase.

叶酸分子是由喋呤和对氨基苯甲酸从头合成的，二者合成的关键酶腺苷三磷酸环化水解酶和氨基脱氧分支酸合成酶已得到克隆鉴定。

In this study, we performed sequential combined PI and DAPI staining and FISH with two different 45S rDNA clones on meiotic pachytene and mitotic metaphase chromosomes in tomato.

采用CPD染色对番茄减数分裂粗线期和有丝分裂中期染色体进行了显带分析，随后用两种不同的45S rDNA 克隆在相同的分裂相进行了荧光原位杂交定位分析。

FISH with the tomato 45S rDNA clone revealed very strong signal in the satellite on the short arm of chromosome 2 as well as weak signals in five CPD banded regions at pachytene or four pairs of CPD banded regions at metaphase.

用番茄的一个45S rDNA 克隆进行的荧光原位杂交，不仅在位于2号染色体短臂的随体上显示了强的杂交信号，而且在粗线期染色体的5个CPD带区或有丝分裂中期染色体的4对CPD带区显示了弱的杂交信号。

After linearisation by restriction enzyme PacI, the recombinant plasmids were transfected into HEK 293-cells and amplified to give large quantities of infectious recombinant adenovirus carrying the sequences of si-FOXO1s, designated Ad-si-FOXO1-1, Ad-si-FOXO1-2, Ad-si-FOXO1-3, Ad-si-FOXO-pan and Ad-NC, respectively.

人工合成si-FOXO1s的cDNA序列，退火形成的双链cDNA克隆到腺病毒表达载体pAdTrack-CMV上，经PmeI线性化后与pAdEasy-1细胞内同源重组，最后在HEK293细胞中包装为重组腺病毒，分别命名为Ad-si-FOXO1-1、Ad-si-FOXO1-2、Ad-si-FOXO1-3、Ad-si-FOXO-pan和Ad-NC。

TNFR1/RANK chimera cloning of cDNA, with its plat E-retroviral package by retroviral infection such transfer to TNFR1/R2 gene knock-out mice (TNFR1-/R2-) Bone marrow macrophages in; TNFɑ used to stimulate, respectively, to observe the OC and differentiation of bone re-absorption capacity; confirmed by Western Blot method TNFR1/RANK through NF-?

克隆TNFR1/RANK嵌合体的cDNA，用plat E将其包装成逆转录病毒，通过感染将这种逆转录病毒转染到敲除TNFR1/R2基因的小鼠(TNFR1-/R2-)骨髓巨噬细胞中；用TNFɑ刺激，分别观察OC的分化及骨的重吸收能力；用Western Blot方法证实TNFR1/RANK是通过NF-？

The expression of CD59 on the cell membrane was tested by cell immunohistochemistry and flow cytometry. Results pALTER plasmid containing CD59 was cut with restriction enzymes and a 496 bp fragment obtained by electrophoresis, which was complete conformity with the insert.

将两种含有不同突变体的人CD59全长cDNA序列重组pALTER质粒，应用阳离子脂质体导入法与pcDNA共转染CHO细胞，用G418筛选阳性克隆，应用细胞免疫组化及流式细胞仪检测CD59在CHO细胞表面的表达。

Methods: Two recombinant palter plasmid containing whole CDNA sequences of HMCD59 transfected CHO cells by lipofectamine with PCDNA respectively. Then G418 was used to select positive clone.

分别构建两种含有HMCD59全长CDNA序列的重组pALTER质粒，并与pCDNA3质粒，按3：1比例混合后，运用脂质体介导法共转染CHO细胞(编号为HM5-CHO及HM6-CHO)，用新霉素类似物G418初步筛选出阳性克隆。

RESULTS A eukaryotic expression system for high expression humanmutantCD59 were successfully set up : The recombinant PALTER-MAX plasmid containing human mutantCD59 cDNA and PCDNA plasmid were co-transfected into CHO cell by cation lipoid mediating method ;and the cells were grown in F12 medium containing 400ug/ml G418 for 14 days, positive clones were grown in RPMI1640 medium to get stable expressing cell lines . Highly expressing clones were selected by flow cytometry ,and were named PALTER-CD59-CHO1PALTER-CD59-CHO2 . Flow cytometry indicated that expression rates of PALTER-CD59-CHO1 and PALTER-CD59-CHO2 were 53.7%and 54.5%. Further more, Stable highly expressing CHO cell lines were more detected by immunocytochemistry and immunofluorescence technology . PALTER-CD59 -CHO1 and PALTER-CD59-CHO2 were grown in RPMI1640 to get a large of cells . CD59 protein were obtained by spalling PALTER- CD59- CHO1 and PALTER - CD59 - CHO2 cells . Stable highly expressing cells were further validated by SDS-PAGE, immunoblot analysis and solid enzyme immunoassay . PALTER - CD59 - CHO1 and PALTER - CD59 - CHO2 were glycated in RPMI1640 of 50mM ribose for 72 hours .BCECF releasing test indicted that the releasing rate of PALTER-CD59-CHO1 or PALTER-CD59-CHO2 was less high than PALTER-CHO ,and the releasing rate of glycated PALTER-CD59 -CHOI or PALTER-CD59-CHO2 was higher than unglycated ones . PALTER -CD59-CHO1 and PALTER -CD59 -CHO2 were glycated in RPMI1640 of 50mM ribose for 72 hours .BCECF releasing test indicted that the releasing rate of PALTER - CD59 - CHOI or PALTER - CD59 - CHO2 was less high than PALTER-CHO ,and the releasing rate of glycated PALTER-CD59-CHO1 or PALTER - CD5 9-CHO2 was higher than unglycated ones .

结果 成功构建突变人CD59的真核细胞表达系统：运用阳离子脂质体介导法将含有突变人CD59的PALTER—MAX重组质粒与PCDNA共转染入CHO细胞：用含有400ug／mlG418的F12培养基培养14天，筛选出稳定阳性表达克隆，RPMI1640培养基扩增获得稳定表达细胞株，并用流式细胞术进一步筛选出高效表达细胞株分别命名为PALTER—CD59—CH01、PALTER—CD59—CH02，表达率分别为53.7％、54.5％；应用免疫组化方法、免疫荧光技术进一步鉴定阳性细胞株；RPMI1640培养基大量扩增PALTER—CD59—CH01、PALTER—CD59—CH02细胞株，裂解细胞得到CD59蛋白质；通过SDS—PAGE凝胶电泳技术、免疫印迹技术、固相酶联免疫吸附试验验证了这两中文摘要个阳性细胞株CO59蛋白的高效表达；50mM核糖培养72小时，获得突变人CD59糖化细胞株，BCECF染料释放试验结果显示，PALTER一CD59一CHOI、pALTER一CD59一CHOZ细胞较PALTER一CHO细胞染料释放率低，未糖化PALTER一CD59一CHOI、PALTER一CD59一CHOZ细胞比较糖化后细胞染料释放率低。

Panhandle PCR is a quick and convenient new technique for cloning and identifying unknown partner genes.

这种特殊的PCR方法可以高度有效地克隆和鉴定已知片段两侧的未知基因片段。

It has been put forward that there exists single Ball point and double Ball points on the symmetrical connecting-rod curves of equilateral mechanisms.

从鲍尔点的形成原理出发，分析对称连杆曲线上鲍尔点的产生条件，提出等边机构的对称连杆曲线上有单鲍尔点和双鲍尔点。

The factory affiliated to the Group primarily manufactures multiple-purpose pincers, baking kits, knives, scissors, kitchenware, gardening tools and beauty care kits as well as other hardware tools, the annual production value of which reaches US$ 30 million dollars.

集团所属工厂主要生产多用钳、烤具、刀具、剪刀、厨具、花园工具、美容套等五金产品，年生产总值3000万美元，产品价廉物美、选料上乘、质量保证，深受国内外客户的青睐