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GFP gene eukaryotic express plasmids were transfected into ES cells. The cells were observed under the inverted fluorescent microscope. The clones which expressed the powerful green fluorescence were chosen to be named ES-GFP cells and cultured continuously. The examination of ES cell totipotent including Alkine phosphatase staining, embryonic body formation in vitro and teratomas formation in nude mice was implemented.

1。采用脂质体方法将GFP质粒转染ES-D3细胞株，经筛选后倒置荧光显微镜观察细胞形态及荧光状态，挑取荧光最强的克隆，命名为ES-GFP细胞，进一步扩增培养，观察细胞生长及荧光表达情况，行细胞分化全能性鉴定包括碱性磷酸酶（alkine Phosphtase，AKP）染色、体外胚胎体（embryonic body，EB）形成实验及裸鼠体内成瘤实验。2。

Cloning of the Schwanniomyces occidentalis α-amylase and high expression in S.cerevisiae.The E.coli / yeast shuttle plasmid YCEpl partial library of Schwanniomyces occidentalis DNA was constructed and α-amylase gene fragments were screened in Saccharomyces cerevisiae by amylolytic activity.Several transformants with amylolysis were obtained and one of the fusion plasmids has about 5.0 kb inserted DNA fragment.It contains the upstream and downstream sequences of α-amylase gene from S.occidentalis .

以E.coli/yeast穿梭质粒YCEp1为载体构建西方许旺酵母部分基因组文库，在大肠杆菌中扩增后提取混合质粒DNA，经电转化非缺陷标志酒精酵母 AS.2.1364，在YPDS平板(含1%葡萄糖和1%可溶性淀粉)上用淀粉水解活力筛选含水解淀粉的阳性转化子，从阳性转化子中分离重组质粒证实含5.0kb的插入片段，用α-淀粉酶基因两端序列设计的引物PCR扩增及扩增片段序列分析证实该片段中含有α-淀粉酶全部编码序列。

Instead of plasmids the strong bacteriophage T5 promoter has been used to replace native promoters in E.

而非质粒强烈噬菌体T5的启动子已被用来取代本土启动子在大肠杆菌中。

Japonicum was firstly PCR amplified from S. japonicum cercaria cDNA library. Its recombinant prokaryotic and eukaryotic expression plasmids were successfully constructed and its nucleotide sequence was determined. It was firstly expressed in E. coli with MW of about 17 ku.

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In the works,DNAssist and ANTHEPROT V5 were used to analyze the sequence,the secondary structure,and the physical and chymic character.Results The inserted fragments in all constructed plasmids were structurally confirmed to be consistent with that of the published data.

在构建融合蛋白之后，运用DNA分析软件DNAssist和蛋白质分析软件ANTHEPROT V5分析融合蛋白的氨基酸序列、二级结构及其理化性质。

To construct recombinant adenovirus,the N and G genes were respectively cleaved from the pMD18-T plasmids by double restriction endonuclease,and then cloned into shuttle vector pAdTrack-CMV coinstantaneously.

经卡那霉素抗性和酶切筛选，最终成功构建了含有N基因和G基因的重组穿梭载体pAdTrack-CMV-N-G。

In HeLa cells trasnfected with GFP-BGLF4 expressing plasmids, cells were sorted into two groups with various levels of BGLF4 expression by FACS. We found that these two populations of cells display dramatically different cell cycle profile.

若利用HeLa细胞将GFP-BGLF4转染送入细胞后，对BGLF4表现量多或表现量少的细胞群各别分析其DNA含量，则发现BGLF4表现量高时S-phase细胞明显增加，而且这些细胞的细胞周期将无法离开S-phase往G2/M-phase过渡转换时期推进。

METHODS: Tum5 gene was amplified from plasmid pSPORT1Sfi by PCR technique and subcloned into the expression plasmid of lentiviral vector, pGCFU, to generate the lentiviral expression vector, pGCFUTum5. The correct Tum5 gene was confirmed by endoenzyme digestion and sequencing. Recombinant lentiviruses were produced by 293T cells following the cotransfection of pGCFU Tum5 and packaging plasmids-pHelper1.0and pHelper2.0. The resulting recombinant lentiviruses (GCFUTum5) which carried Tum5 and EGFPgene were then used to infect human umbilical vein endothelial cells.

采用PCR技术从含有tumstatin基因的质粒克隆模板 pSPORT1Sfi钓取Tum5基因，并将基因克隆到慢病毒载体表达质粒pGCFU中，构建慢病毒载体表达质粒pGCFUTum5，通过酶切、测序验证Tum5基因后，将pGCFUTum5质粒和包装质粒pHelper1.0，pHelper2.0共同转染人胚胎肾上皮细胞系293T细胞，获得携带Tum5基因和EGFP基因的重组慢病毒GCFUTum5，并转染靶细胞人脐静脉血管内皮细胞。

RESULTS:①Enzymolysis and nucleotide sequence analysis of recombinant plasmids showed that two VEGF-C siRNA eukaryotic expression vectors were constructed successfully.

结果：①重组质粒的酶切、核苷酸序列分析结果：经酶切和测序鉴定证实成功构建了血管内皮生长因子C小干扰RNA表达载体，建立了稳定转染细胞株。

The stability of exogenous plasmids introduced in BMB171 was correlated with the patterns of replicons, and sizes of plasmids.

用pHT3101、pBMB3305、pBMB1736、pBMB671、pBTL-1和pHV1249等6种外源质粒电转化无质粒突变株BMB171的转化频率，分别是Bt-4Q7、Bt-4D10和Bti。

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！