质粒

They harbor indigenously circular as well as linear plasmids, a useful source for developing genetic tool for studying of their unique physiology and metabolism.

在许多稀有放线菌发现了环型与线型质粒，由质粒发展起来的遗传操作系统可以为研究稀有放线菌的生理及代谢提供强有力的工具。

Part two, The consructon of universal probe real time PCR detecting of Avian viral disease We constructed a plasmid including a specified fragment of NDV, and optimizated reactive condition of universal probe real time PCR, and detected Infectious Laryngotracheitis Viruse, Infectious Bronchitis Viruse and

第二部分：通用探针实时荧光PCR检测禽病毒病方法的建立本部分首先构建含有NDV特定片段的质粒，以提取的质粒为模板

So it is assumed that the E-box might be associated with the enhancement of transcription activity.Here, the LTRs of WH17 ,DLA-25,DLA-118 and LTR with single mutation in the E-box motif were separately cloned into pCAT-Basic vector resulting in a series of recombinant plasmids containing CAT reporter under the control of different versions of LTRs.

通过对EIAV分离株WH17的LTR与L株、DA和DLA株的LTR序列进行比较，发现其在在U3-R结合处多一个E-box基序，推测该基序的变化可能会起到促进转录作用，为此将EIAV强毒株(DLA-25)、驴白细胞弱毒株(DLA-118)、EIAV分离株WH17以及U3-R结合处的E-box基序点突变的LTR分别克隆到pCAT3-basic质粒中的报告基因氯霉素乙酰转移酶的上游，获得一系列受不同LTR控制的CAT表达质粒。

Its cDNA had been cloned by RT-PCR. The results of comparison on sequence homology suggested that Luffin P1 may be derived from a vicilin like precursor which is a storage protein in the seeds of Luffa cylindrica.

本文利用3'RACE法克隆了它的cDNA，并构建了它的融合表达质粒pGEX-4T-S2，该质粒在大肠杆菌中获得了成功表达。

Methods:The luciferase reporter plasmids of MDM2 P2 promoter containing different SNP309 were contructed,and cotransfected into CV-1 cells with AML1-ETO,AML1 and SP1 expression plasmid.The transactivities of luciferase were assayed by luminometer to determine the impact of AML1,AML1-ETO,and SP1 on MDM2 P2 promoter.

研究方法：构建含不同SNP309位点的MDM2 P2启动子的报告质粒，与AML1-ETO、AML1以及SP1的表达质粒单独或共转染CV-1细胞，测定荧光素酶的活性，分析AML1-ETO、AML1以及SP1对MDM2 P2启动子的影响。

Methods:The expressive plasmids of AML1b and AML1-ETO were transfected into CV-1 and 293 cells.The expression level of endogenous pig7 gene was detected by Realtime-PCR.The luciferase reporter plasmids containing pig7 enhancer/corresponding mutant sequences were constructed and co-transfected into CV-1 cells with expressive plasmids of AML1b and AML1-ETO.The transactivity of pig7 enhancer was assayed by luminometer.

研究方法：AML1b和AML7-ETO表达质粒分别转染及共转染CV-1和293细胞，用Realtime-PCR方法检测上述基因对两种细胞内源性pig7基因mRNA表达水平的影响；构建含AML1b结合位点的pig7基因增强子序列以及相对应的突变位点的荧光素酶报告基因质粒；将表达载体/突变载体与AML1b或AML1-ETO基因瞬时共转染CV-1细胞，分析AML1b、AML1-ETO对报告基因的转录调节作用。

To prove that the cloned DNA fragment can express tryptopanase,a new plasmid pET28C-TnaA , in which the cloned DNA fragment was located downstream of T7 promoter on pET28c was constructed and transformed into host BL21(DE3),a BL21 lysogen of bacteriophage DE3 in which the only promoter known to direct transcription of the T7 RNA polymerase gene is the lacuvS promoter ,which is inducible by IPTG.

为了证明质粒上的基因能表达出有活性的色氨酸酶，将这个DNA片段克隆到PET28c质粒的BamHI和HindⅢ位点上，使该片段受T7 RNA聚合酶的启动子控制，然后转化噬菌体DE3的溶源菌BL21(DE3)。

Coli XL-1 strain, purposed cassette recombination plasmids pNB0098-K and pNB0097-K were constructed, in which the purposed gene was linked with plasmid pUC18, and the marker gene kanamycin was inserted in the purposed gene. The linear cassette DNA fragment was mixed in the medium of cultured competent cells of meningococcus BT878. The function of the purposed gene was inhibited by the homologous replace of casstte gene.

将线性的重组质粒片段置于培养基中，与感受态的脑膜炎双球菌混合培养，重组质粒中的同源性片段被脑膜炎双球菌细胞识别和摄取，整合到染色体上，目的基因被带有卡那霉素基因插入的同源性重组片段代替，导致失活，在选择性培养基上筛选出基因被敲除的突变株。

The nucleoprotein gene and glycoprotein gene of rabies virus were amplified by RT-PCR and then cloned into pMD18-T plasmid,respectively.

将N基因和G基因分别克隆入质粒载体pMD18-T后，对筛选的含有N基因和G基因的重组质粒进行了全基因序列测定和分析。

In our experiment, the GADes gene from rat brain was cloned and expressed in E.coli, so as to make the recombinant GADes produced by gene technology for clinical application and further study on the relation between GAD and type 1 diabetes mellitus.Firstly, with the pUClS/GADes recombinant plasmid as template, the full-length encoding sequence of GADes was amplified by PCR, and cloned into pGEM-T vector. Secondly, the target gene was cut by EcoRI and inserted into restriction sites of pET42a vector for expression.

本研究首先用PCR方法，以pUC18/GAD_(65)为模板，扩增目的基因，克隆到pGEM-T载体中，构建pGEM-T/GAD_(65)重组质粒，然后用EcoRI酶切，切下的目的基因片段插入pET42a融合型表达载体中，经酶切鉴定和测序，证实插入方向、读码框架正确；重组表达质粒转化大肠杆菌BL21(DE_3)中诱导表达，表达产物经亲和层析纯化后获得了融合蛋白GST-GAD_(65)。

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！