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MAIN OUTCOME MEASURES: The results of recombinant plasmid PCR and gene sequencing, the correlation coefficient of the standard curve and the repeatability of the plasmid standard.

主要观察指标：重组质粒聚合酶链反应和基因测序结果，标准品标准曲线的相关系数及标准品的重复性。

A PCR product was amplified using RT-PCR method from total RNA from the abdomens of R strain using the degerated primer. The PCR product was purificated and cloned. The plasmid is pBluescript Ⅱ KS. Recombined plasmid DNA was identified by endoenzyme. The results indicated there were different P450 genes in the PCR product.

运用这对简并引物首次从德国小蠊抗拟除虫菊酯的品系R中扩增出的特异性的DNA片段，经纯化、连接和转化，重组至pBS质粒，筛选阳性克隆，提取重组质粒的DNA，并经初步的酶切鉴定，发现存在着不同的CYP4基因。

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。

In this study, a recombinant plasmid generating short hairpin RNA was constructed — To construct the recombinant plasmid carrying shRNA to HPV6bL1 and analyze the nucleicacid sequence for further searching new gene therapy method of cauliflower excrescence.

目前对于尖锐湿疣仍无有效的病毒特异性防治措施来保护人类免受其危害，而基因疫苗则为预防HPV感染提供了可能性；L1是HPV6型主要的衣壳蛋白，具有较强的免疫原性，可刺激机体产生保护性的中和抗体。

The flic gene of App was obtained by PCR based on highly homology with Escherichia coli, Salmonella, Shigellosis. Then it was legated to pMD-18T vector. The recombinant plasmid was identified by enzyme and sequence analysis. The 528 bp fragment of flic gene was successfully cloned into pGEX-KG vector, then the recombinant plasmid was transformed into E.

以猪传染性胸膜肺炎放线杆菌血清Ⅰ型菌株基因组为模板，根据其鞭毛区与其它细菌高度同源性设计引物，利用PCR方法扩增鞭毛蛋白基因flic片段，将其亚克隆到pMD-18T载体中并进行PCR和酶切鉴定，再将其亚克隆与载体pGEx-kG分别双酶切和连接，构建重组表达载体pGEX-flic，并将其转化大肠杆菌工程菌BL21，进行原核表达。

It is the fact that three methionines distributed in the Hsp16.3 will begradually oxidated, in vitro, by hydrogen peroxide(H2O2). When the hsp16. 3 gene was cloned with pMV261 plasmid vector and transfected into Mycobacterium smegmatis, we found that the ability of anti-H2O2 will be decreased greatly among the strains of M. smeg. containing pMV261-hspl6. 3 while compared with the control strains of M. smeg. containing free plasmid vector. But we found that the ability of anti-H2O2 will be increased greatly among the strains of M. smeg. containing pMV261-38kDa while compared with the same control.

尽管Hsp16.3中有三个甲硫氨酸在体外能被过氧化氢(H_2O_2)逐一氧化，我们将Hsp16.3基因克隆到pMV261质粒上，并转染到耻垢分枝杆菌中发现，转染了pMV261-16.3的耻垢分枝杆菌抗过氧化氢的能力大大下降，而对照含pMV261-38kDa的耻垢分枝杆菌抗过氧化氢的能力却有明显的增强，据此，我们认为，Hsp16.3在细胞内可能不具有抗过氧化氢的能力。

pPR2 is the smallest ac-tinomycete linear plasmid beyond Streptomyces. This is the first time to report the complete nucleotide sequence of linear plasmid in the genus Planobispora. pPR2 may have novel mechanism for telomere replication, and pPR2.2c and pPR2.3c encode the possible telomere replication proteins.

pPR2是链霉菌之外的放线菌中最小的线型质粒，其序列在游动双孢菌属的线型质粒中是首次报道。pPR2可能具有新型的端粒复制的机制，其中pPR2.2c和pPR2.3c编码可能的端粒复制蛋白。

The Cre gene was cloned from lysogen BM25.8 by PCR. The Cre gene was then inserted in between the CaMV35S-35S double promtor and Nos terminator of PBI525 plasmid after sequencing. The expression box was cut down and inserted into plasmid pBINPLUS to obtain the final fertility-restoring expression vector pBINPLUSCre.

PCR方法从溶原菌BM25.8基因组DNA扩增出Cre基因，克隆测序验证无误后插入PBI525 CaMV35S-35S双启动子和Nos终止子之间，再将表达盒切下插入pBINPLUS质粒相应位点，得到恢复基因植物表达载体pBINPLUSCre。

BALB/c mice were co-inoculated muscularly by this plasmid and the nucleic acid vaccine plasmid pVAXGE expressing the HIV-1 (Human immunodeficiency virus type I) gag-gp120 protein. The level of serum antibodies after immunization showed that the specific antibodies against HIV-1 appeared at the second week and raised to the peak level at the sixth week for the co-immunization group.

将它与表达I型人免疫缺陷病毒(Human immunodeficiency virus 1， HIV-1) gag-gp120的核酸疫苗质粒pVAXGE共同肌肉注射BALB/c小鼠，免疫3次后，以ELISA法检测免疫小鼠血清中抗HIV-1抗体水平，结果显示联合免疫组小鼠在免疫2周后已有抗体产生，6周后进入高峰。

Pocine interferon α mature protein genes were amplified by reverse transcription polymerase chain reaction, and the recombinant replication-defective human adenovius serotype 5 plasmid pAd-poIFN-α was constructed. When the recombinant plasmid pAd-poIFN-α was linearized with PacI, and then transferred into HEK-293A cells, the virus plaque was isolated and purified in HEK-293A cells by three of plaque purification passages.

本研究利用RT-PCR方法扩增猪α干扰素成熟蛋白基因后构建了重组腺病毒质粒pAd-poIFN-α，经PacI酶切后转染HEK-293A细胞，3次噬斑纯化后获得了重组腺病毒rAd-poIFN-α。

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！