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To construct a plasmid for analyzing gene functions and expressions and to study the MXAN1334 gene in Myxococcus xanthus with the plasmid.

旨在构建一个用于基因插入失活，并同时可通过报告基因分析插入位点基因表达情况的质粒载体，并应用该质粒对黄色粘球菌MXAN1334基因功能和表达情况进行分析。

Methods: The human KDR promoter (-225 bp~+127 bp) was cloned by PCR. Subsequently, a recombinant adenoviral plasmid carrying KDR promoter for the HSV-tk gene was constructed with the AdEasy system. As a control, the plasmid pAdCMV-tk carrying CMV promoter for the HSV-tk gene was also constructed. 293 packaging cells were transfected by both newly-constructed plasmids and the infectious viruses AdKDR-tk and AdCMV-tk were generated. Then the KDR-producing cells and the KDR nonproducing cells (HepG2) were infected with AdKDR-tkand AdCMV-tk, followed by ganciclovir administration.

应用PCR克隆出人KDR基因启动子序列－225bp~+127bp，以AdEasy system为载体，构建携带受KDR启动子或CMV启动子调控tk基因表达的2种重组腺病毒质粒pAdKDR-tk和pAdCMV-tk，在293细胞中包装、扩增后，体外感染表达KDR的脐静脉血管内皮细胞系HUVEC和不表达KDR的肝癌细胞系HepG2，并给予不同浓度的丙氧鸟苷处理，5d后收集存活细胞并计数。

Objective:To recombine the plasmid PcDNA3.1-sTNFRⅠ,and detect the expression of recombinant plasmid in pericementum cell,and evaluate the bioactivity of expressed sTNFRⅠ.

目的：构建真核表达质粒PcDNA3.1-sTNFRⅠ，并转染人牙周膜细胞观察是否有目的产物的表达，研究其是否具有生物学活性，为进一步研究其在牙周治疗中的作用提供实验基础。

Objective To investigate the possibility of self-assembling gene nanoparticles between the amphiphilic chitosan derivative and plasmid DNA, and study the interaction mechanism between N -methylene phosphonic chitosan and plasmid DNA, which might provide a new strategy for developing safe and efficient non-viral vectors.

目的 探讨双亲性壳聚糖衍生物与质粒DNA通过自组装的方式形成基因纳米粒子的可行性，考察载体与质粒DNA的相互作用机制，为开发安全高效的非病毒载体提供新的途径。

The phytopathogenic fungus Fusarium graminearum has been transformedusing a plasmid (pAN7-1) containing the Escherichia coli hygromycinphosphotransferase gene. Stable hygromycin-resistant transformant coloniesappeared at frequencies between 2 and 10 per μg plasmid DNA.

将含有大肠杆菌中潮霉素B磷酸转移酶控制基因的质粒pAN7-1转化入小麦赤霉病菌野生菌株中，结果表明，pAN7-1的转化效率为2-10个/μg质粒DNA。

The fusion gene was inserted downstream to the MOX promoter of Hansenula polymorpha shuttle plasmid pDGXHP2.0, and the constructed multicopy secretory recombinant plasmid was transformed to Hansenula polymorpha ATCC34438 Ura3(superscript - by electroporation.

将融合基因插入汉逊酵母穿梭质粒pDGXHP2.0的甲醇氧化酶基因启动子下游，构建成多拷贝分泌型重组表达载体。

Through the test of retransformation with plasmid pUB110 fromtransformation with plasmid pUB110 from transferant and passage test,B.licheniformis 6104 and its mutant 610401 were proved to be a good receptorfor exogenous genes,especially for lysine synthetase expression.

通过质粒的再转化试验及传代稳定性试验，进一步证实B.licheniformis 6104及其突变菌株610401是较好的受体菌，尤其是用于赖氨酸合成酶基因的表达。

The plasmid pAO815 is then used in homologous recombination, and the recombinant plasmid pETnsGH is further converted, jointed and inserted into GS115 saccharomycetes for the foreign gene to express normally.

再选择质粒pAO815进行同源重组，进而有效地将重组质粒pETnsGH转化接合插入GS115菌种中，使外源基因sGH正常表达。

Methods Amplify IL-6 (23)-PE40 gene from the previously constructed recombinant plasmid pKK-IL-6(23)-PE40 by PCR, and insert to the SnaB Ⅰand NotⅠ sites of vector pPIC9. Transform the constructed recombinant plasmid pPIC0-SN to P. pastoris GS115 by electroporation, screen Mut-recombinants for expression under induction of methanol.

采用PCR技术，将目的基因IL-6(23)-PE40插入到pPIC9载体中SnaBⅠ与NotⅠ位点，电转化至巴斯德毕赤酵母GS115中，筛选Mut-型重组酵母；甲醇诱导表达，体外细胞试验检测其细胞毒性。

Methods The hTERT promoter as E1A and E1Bp gene were amplified by PCR; the GFP gene and GM-CSF were also amplified by PCR, and the products were subcloned into Teasy plasmid in certain order to generate pSh-GFP-SV55K plasmid.

采用PCR的方法克隆腺病毒E1区基因(E1A， E1B55K)、EIB的启动子(E1Bp)以及人端粒酶逆转录酶亚基启动子，以及2个目的基因：人GM-CSF和GFP(Green Fluorescence Protein， GFP)。

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！