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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。

The key to this riddle appeal to be plasmids, which can block phage adsorption.

这个难题的关键是质粒，它能够阻碍噬菌体的吸附作用。

Locations of transgenes in the recovered plasmids were determined by southern blotting. And then adjacent sequences of transgenes integration sites in three recovered aberrant classes were subcloned and analyzed. Transgene of A-2 was inserted into common carp DNA sequences homologous to the mouse phosphoglycerate kinase-1 gene and those of A-3 and A-6 were inserted into common carp DNA sequences homologous to the human epidermal keratin 14 gene and sequence of common carp β-actin gene, respectively.

通过Southern杂交对转植基因在回收质粒上的位置进行了定位分析，并对三种回收到的变异型转植基因旁侧顺序进行了亚克隆和顺序测定。A-2的转植基因整合到了与小鼠磷酸甘油酸盐激酶-1基因同源的鲤鱼基因组顺序旁侧，A-3的转植基因整合到了与人表皮角蛋白14基因同源的鲤鱼基因组顺序旁侧，A-6的转植基因整合到了普通鲤鱼β-actin基因顺序中。

However, there are still several safety concerns about the use of a DNA vaccine, which include the possibility of integration into the host genome, adverse immunopathology, and anti-DNA autoantibody induction.Attempting to solve some of these problems, eukaryotic expression plasmids pTL-8/ AMA-1 and pTL-8/ AMA-1 which express trans-activator or reverse trans-activator, respectively, and AMA-1 gene of Plasmodium falciparum were constructed by using tetracycline regulable system.

首先构建了恶性疟原虫顶第四军医大学硕士学位论文端膜抗原1(AMA一1)基因和反式活化因子基因的真核表达质粒pTL一8/灿认一l和PTL一8/灿认一1，并大量制备这两种质粒及表达转录抑制子的质粒pUHS6一1。

Methods Six recombinant plasmids with shRNAs targeting the PDE5A3 gene of Homo sapien were constructed.

方法构建6个靶向人PDEsA3基因的短发夹RNA重组质粒，转染人阴茎海绵体平滑肌细胞48 h后，逆转录-聚合酶链反应及Western blot检测PDE5A3基因的表达抑制效果。

In this study,two recombinant plasmids harboring non-structural and structural protein gene named pMD18-T-NS1 and pMD18-T-NS1 were constructed and sequenced.

但这种诊断试剂和亚单位疫苗的研制，必须建立在对病毒蛋白抗原性的研究之上。

Expression of M, NP, F and HN genes were detected and confirmed by the indirect immunofluorescence analysis. The syncytium formation was observed in the HeLa cells co-transfected with pCAGG-M, pCAGG-NP, pCAGG-F and pCAGG-HN recombinant plasmids at 48h post transfection.

纯化后的重组质粒通过脂质体单独转染HeLa细胞，经间接免疫荧光试验检测到了M、NP、F和HN蛋白的表达。pCAGG-M、 pCAGG-NP、pCAGG-F和pCAGG-HN重组质粒组合共转染HeLa细胞48h后，可以观察到明显的细胞融合现象。

Results The three plasmids were effectively cotransfected and a high titre of lentivi-rus was obtained (2×10 6 IU/ml).

结果共转染293T细胞后获得较高滴度的慢病毒(2×106IU/ml)。

METHODS: MyoD cDNA fragments were extracted from plasmids pEMSV-MyoD with polymerase chain reaction, and PCR was used to clone the whole-length gene of MyoD. After adding CACC sequence at 5' end, MyoD gene was cloned by orient topology into transfer ventor, pENTR/D-TOPO. Objective gene was transferred into adenoviral expression vector DNA via pENTR/D-TOPO vector. The recombinant adenoviral vectors transfected into HEK293A cells by using lipofectamine were packaged and amplified.

从pEMSV-MyoD质粒上用聚合酶链反应法扩增出MyoD cDNA片段，再通过聚合酶链反应使MyoD基因加上CACC序列接头，经过定向拓扑克隆使目的基因连接到转移载体上，再通过LR酶促反应，将目的基因转移到腺病毒表达载体DNA上，获得MyoD基因重组的腺病毒DNA，用脂质体转染法转染HEK293A细胞，包装扩增出MyoD基因重组的腺病毒。

This method of primer design can be used to construct mutant plasmids with great ease and speed.

这种方法能用于超过20bp的碱基的插入或替代突变，及用于超过2000bp DNA片段的删除突变的研究。

She gently rebuff ed him, but agreed that they could be friends

她婉言拒绝了，但同意作为朋友相处。

If in the penal farm, you were sure to be criticized.

要是在劳改农场，你等着挨绳子吧！