质粒

The RT-PCR product was inserted into pTG19-T vector and transformed into E. coli successfully. By blastn, the sequence results of Kunming mus musculus were in complete accordance with the conservative sequence of Genbank NR_003278 (791bp-1153bp). By Blastn in NCBI, the sequence with little difference among animals was confirmed to be conservative. After Blastn, fourteen complete CDS coding for different animals were chosen. According to VECTOR NIT 9.0 software, the similarities between Kunming mus musculus and bos taurus, homo sapiens, erinaceus europaeus, cricetulus griseus, sus scrofa, dasypus novemcinctus, rattus norvegicus, rabbit, equus caballus, macaca fascicularis, didelphis virginiana, monodelphis domestica and vombatus ursinus was 67%, 100%, 100%, 36%, 100%, 100%, 67%, 100%, 100%, 92%, 99%, 99% and 99%. In the phylogenetic tress constructed with the forteen 18S rRNA by Treeview, the Kunming mus musculus clustered with cricetulus griseus, sus scrofa and rabbit, which was nearer to cricetulus griseus and was most far away from macaca fascicularis.(3) After sencodary structure analyses of 18S rRNA of mus musculus, an oligonucleotide fragment for RNAi was designed and synthesized, which was transformed into plasmid, and restriction enzyme analyses and sequencing results should the expression plasmid pGPH1/ GFP/Neo-mouse-sh 18S rRNA were constructed for RNAi successfully.

结果①通过RT-PCR检测显示18S rRNA基因在小鼠卵巢组织和单个GV期、MⅠ期卵母细胞中均有表达，且在未成熟卵母细胞中，MⅠ期的表达明显强于GV期的表达；②RT-PCR产物克隆测序结果显示：昆明小鼠18S rRNA基因保守区序列与基因库序列[NR_003278保守区部分(791bp～1153bp)]完全一致；Blastn比对结果发现：在不同物种中差异较小，选出14种生物18S rRNA全序列经VECTOR NIT 9.0软件分析，提示昆明小鼠18SrRNA与牛、人类、刺猬、中国仓鼠、猪、犰狳、褐鼠、兔子、马、食蟹猴、负鼠、短尾猊、袋熊的18S rRNA的相似率依次为67%，100%，100%，36%，100%，100%，67%，100%，100%，92%，99%，99%，99%；Clustal 1.81和Treeview构建出的分子进化树表明：在上述14种生物中昆明小鼠与中国仓鼠进化关系最近，与兔子、猪聚成一簇，与食蟹猴进化关系最远；③根据18S rRNA二级结构设计并合成RNA干扰寡核苷酸片段，重组质粒经过限制性内切酶及测序表明成功构建了pGPH1/GFP/Neo-mouse-sh 18S rRNA干扰表达质粒。

Methods The plasmid pCEP4P53 expressing human p53 protein and the plasmid pCEP4ASCMH expressing human antisense c - myc gene in eukaryocyte were co - transfected to human bladder cancer cell T24 strain using FuGENE6 as a media, and the results were analyzed by MTT method, cell colony - forming test in soft agar medium and agarose gel electrophore...

将可在真核细胞表达P53蛋白的质粒pCEP4P53和表达反义c-myc基因的质粒pGEP4ASCMHC以染试剂Fu-GENE6为介导，同时转染到入膀胱癌细胞T24中，经MTT检测、软琼脂集落生长、琼脂糖电泳等方法分析实验结果。

In this paper, an escherichia coli-saccharopolyspora erythraea shuttle vector containing the erya promoter region was constructed using the enhanced green fluorescent protein gene as a reporter. the shuttle plasmid was transformed into sac.erythraea a226 and streptomyces lividans jt46 by peg mediation, respectively. fluorescence microscopy confirmed that the egfp was expressed in both strains.

本文克隆了erya基因的启动子perya，以绿色荧光蛋白基因为报告基因，构建了大肠埃希菌-糖多孢红霉菌穿梭型质粒。peg介导原生质体转化法将穿梭型质粒分别转入糖多孢红霉菌a226与变铅青链霉菌jt46，荧光显微镜检测发现，此启动子在两菌株中都具有功能。

As most Streptomyces promoters are invalid in S. erythraea, the promoter of erythromycin-resistant gene, ermE, and Streptomyces chromosomic integration site, attP, and apramycin-resistant gene, apr from plasmid pSET152 are utilized to construct the expression vector pZMW.

因大多数链霉菌质粒的启动子在糖多孢红霉菌中都不能发挥作用，故利用糖多孢红霉菌染色体上红霉素抗性基因PermE启动子作表达载体启动子，并利用pSET152质粒上链霉菌染色体整合位点和氨朴霉素抗性基因，构建了表达载体pZMW。

Was used to cons Methods: The pSUPER plasmid cloned into RNA polymeraseⅢ H1 promoter was used to construct new vectors which can code hereditable shRNA aimed to HPV18E6/E7 gene mRNA.

采用克隆有人Ⅲ型RNA聚合酶H1启动子的pSUPER质粒，设计并构建针对HPV18E6、E7基因mRNA的、可稳定遗传的编码发夹状结构的特异性RNA干扰质粒载体。

The recombinant plasmids, pIRES-Jurkat Vβ8 and pIRES-Molt4 Vβ2①/②, containing idiotypic TCR Vβ8 frgment of Jurkat cell strain or idiotypic TCR Vβ2 frgments of the Molt4 cell strain were developed.

构建了Jurkat细胞独特型TCR Vβ8 DNA重组质粒pIRES-Jurkat Vβ8和Molt4细胞独特型TCR Vβ2 DNA重组质粒pIRES-MOLT4 Vβ2①/②。

We construct pCDNA3.1-TfRexpression plasmid successfully, there are higher expression of TfR aftertransient and stable transfection; we get positive cell clone of containingpCDNA3.1-TfR, establish infarctate foundation to empirical study ofmolecular imaging of MR.

成功构建了pCDNA3.1-TfR表达质粒；质粒转染U251细胞后瞬时与稳定表达的TfR蛋白明显增高；成功筛选出pCDNA3.1-TfR阳性细胞克隆，为分子影像学的实验研究奠定了坚实的基础。

Methods By the bioinformatics analyzing tools in bioinformatics webs site,a Ta CRISP 2 fulllength gene from the Taenia saginata asiatica fulllength cDNA plasmid libratory was identified and the coding region sequence and the characteristics of the deduced protein were analyzed.The coding region of Ta CRISP was amplified with PCR,endonuclease digestion and cloned into the prokaryotic expression vector pET30a and then expressed in E.coli BL21 with IPTG induction.

用生物信息学方法从亚洲牛带绦虫成虫全长cDNA质粒文库中识别TaCRISP的全长编码基因并预测编码蛋白质的各种结构与功能；将其编码区序列克隆到原核表达载体pET-30a上，测序鉴定重组质粒，之后进行诱导表达，表达产物经十二烷基磺酸钠-聚丙烯酰胺凝胶电泳鉴定。

Then, we cloned the lipA and its partner gene lipB into the vector pUCPCM and pBBR22b both or separately. Therefore, we got 7 expression plasmids pBBR22blipAB, pBBR22blipA, pUCPCMlipAB, pUCPCMlipA, pUCPCMΔlipAlipB, pUCPCMΔlipA, pUCPCMΔlipB, and then electroporated them into B.

接着把lipA和它的伴侣基因lipB单独或全部克隆到载体pUCPCM和pBBR22b上，构建出pBBR22blipAB、pBBR22blipA、pUCPCMlipAB、pUCPCMlipA、pUCPCMΔlipAlipB、pUCPCMΔlipA、pUCPCMΔlipB七种表达质粒，通过电转化将上述表达质粒转化到含T7 RNAP的B。

Then pMD-P80 is digested by the enzymes Xho I and Apa I, separated and linked to the linear plasmid pEGPF-C1 after digested by Xho I and Apa I. So the recombined expression plasmid pGFP-P80 is achieved. Then it is sequenced, PCR and digested by the restriction enzymes Xho I and Apa I. The results give that pGFP-P80 is achieved successfully and the insert sites, direction and reading frame are all right. It builds the basis for expressing, purifying, gaining p80 protein from mammiferous cells.

将pMD-P80 分别经Xho I 和Apa I 双酶切和回收，然后与经过Xho I 和Apa I 酶解的真核表达载体pEGPF-C1 连接、转化，获得重组质粒，经PCR，Xho I 和Apa I 限制性酶切和序列测定，鉴定为真核表达质粒pEGFP-P80，并且目的基因的插入位置、方向和读码框完全正确，为在哺乳动物细胞中表达并分离纯化p80 蛋白奠定了基础。

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！