质粒体

AIM: To construct recombinant eukaryotic expression plasmid of amastin gene of Leishmania Donovani anddetectexpressionofthegeneinNIH3T3cells.

目的： 构建杜氏利什曼原虫无鞭毛体蛋白编码基因的真核表达重组质粒pcDNA31amastin，并研究其在NIH3T3细胞中的表达。

METHODS: Amastin gene was amplified from nuclear DNA of Leishmania Donovani isolates and cloned into an eukaryotic expression vector pcDNA31.

目的： 构建杜氏利什曼原虫无鞭毛体蛋白编码基因的真核表达重组质粒pcDNA31amastin，并研究其在NIH3T3细胞中的表达。

Finally,alkaline lysis and anion exchange chromatography were used to release and purify plasmids.

研究了包括菌体生长和质粒含量等的关系。

It was demonstrated that the BADHgene promoter region and its two mutants are active in monocot rice calli.

实验表明，质粒pDMC202中GUS基因在BADH基因启动区以及两个启动区突变体的控制下获得了表达。

Part Ⅱ Objective: To study the immunoprotective and the immunotherapic effects of the new nucleic acid vaccine of the hepatitis B: pVAX1-C3d-S2S; respectively in normal and HBV transgenic mice after being inoculated intra- muscularly with this Eukaryotic Expression Plasmid, such as the in situ production of the expressed protein in muscle tissue section and specific humoral immune response in normal mice、the variation of specific antigen、antibody and HBV DNA, especialy the changes of specific antigen in liver.

第二部分新型乙肝核酸疫苗pVAX1 C3d-S〓S免疫预防效应和治疗作用的研究目的：研究含有新型分子佐剂补体C3d的HBV adw血清型的乙肝核酸疫苗重组真核表达质粒pVAX1-C3d-S〓S接种正常BALB／c小鼠和HBV转基因小鼠后，其免疫预防作用（接种局部肌肉中特异性抗原蛋白的表达和所诱生的体液免疫应答规律）以及免疫治疗作用（血清中特异性抗原的表达、特异性抗体的产生、HBV DNA的消长和肝组织中抗原表达的变化）。

The pALTER plasmid was successfully transfected into CHO cells and positive clones were screened and detected immunohistologically.

结果 含有人CD59两种突变体的重组pLATER质粒经酶切和琼脂糖凝胶电泳鉴定，得到长496 bp的电泳条带，与所插入片段完全相符。

Objective To obtain the functional fusion protein of rhoptry protein 2(ROP2) and major surface antigen 1 of Toxoplasma gondii in order to further prepare for the diagnosis and polyvalent vaccines of toxoplasmosis.

目的构建编码弓形虫RH株棒状体蛋白2(ROP2)和主要表面抗原1的重组表达质粒，纯化和复性的融合蛋白为弓形虫病快速诊断试剂盒及蛋白质疫苗的研制作准备。

Pneumoniae FH strain was cloned and the sequence was analysed by M13 DNA sequencing method. Comparing the PCR product sequcence with MP M-129 strain P1 gene, we found that there are 4 bases different. This may result from the different MP DNA templates. The maximum homology is 98.8%. The result confirmed the fidelity and specificity of the amplified target DNA segment by PCP, and suggested that two categories of MP P1 gene still exist a few differences even in the conservation region. The cloning MP DNA segment was labelled by random hexanucleotide priming, after hybridization, the probe detection was completed using an anti-digoxigenin antibody alkaline phosphatase conjugate, and the substrates 5-bromo-4-chloro-3-indolyl phosphate and nitro blue tetrazolium. This hybridization system is much superior to the radioactive probe hybridization, because it is safe, easy to handle and has no limitation of decay time. The time required for colormetic detection is also much less than the corresponding autoradiographic exposure time needed to achieve similar detection limits with 32P-labelled probes. The Dig-probes could be used repeatedly, and this made them not only much convenient to use, but also lower the cost, and worthwhile to be used popularly.

将PCR产物进行重组，并将阳性重组质粒，应用M13测序系统对产物进行DNA序列分析，并与MPM-129株P1基因核苷酸进行同源性比较，发现有4个位置的碱基发生了变化，其同源性为98.8％，证实了PCR所扩增DNA片段的准确性和特异性，同时也证实了不同MP组型的P1基因即使在保守区也存在着一定的差异，将克隆的目的DNA片段用异羟基洋地黄毒苷配基用随机引物法标记制备MP DNA探针，杂交后用碱性磷酸酶标记的抗Dig多克隆抗体与杂交体反应，再用BCIP和NBT呈色，制备MP DNA探针，鉴定所扩增片段的特异性，与同位素探针比较，Dig探针不受半衰期限制，可反复使用，而且价格低廉，值得推广使用。

Immunofluorescence staining shows that, the CaN Aβ protein expression of intrapericardial injection group decreased in some myocytes under epicardium, while other groups show no decreased fluorescence.

三在体心肌CaN AβsiRNA表达质粒转染后，转染各组与心肌肥大模型组CaN AβmRNA水平比较无差异(P＞0.05)，但免疫荧光结果显示心包腔内途径CaN Aβ干扰组较心肌肥大模型组心外膜下部分心肌荧光减弱。

Orange-spotted grouper is a benthic protogynous marine fish with sex inversion. It is one of the most important culturing fish in the southeast of Asia. As in other vertebrates, its growth is mainly controlled by growth hormone . Few data are available concerning the regulation of GH in grouper. In this paper, the regulation of growth hormone release from pituitary of orange-spotted grouper and its mRNA level was determined by radioimmunoassay and RNase protection assay.

应用分子克隆技术，构建了斜带石斑鱼生长激素(Growth hormone，GH)基因重组质粒，在大肠杆菌细胞中获得高效表达，亲和层析纯化斜带石斑鱼GH融合蛋白做为抗原，免疫家兔制备特异性抗体，通过酶联免疫实验、放射免疫测定、离体脑垂体静态孵育和液相杂交／核糖核酸酶保护试验及化学发光检测技术，研究了斜带石斑鱼GH分泌及其mRNA表达的调控机制，对于斜带石斑鱼的生长、繁殖、人工养殖等具有重要意义。

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！