表达的

The results showed that human transferrin could be expressed in these cell lines and mouse or goat mammary gland following transfection, and its expression level could be improved dramatically when it was driven by rabbit enhancer and promotor as compared with goat beta lactoglobulin promotor. Transgenic mice were generated by either microinjection or lentivirus infection.

结果显示所有表达载体均可指导人转铁蛋白在细胞以及小鼠和山羊乳腺中表达，兔转铁蛋白增强子与启动子组合可显著性增加人转铁蛋白的表达水平，且其指导表达的效率明显高于山羊β乳球蛋白启动子。

Transgenic tobacco plants were obtained through screening with kanamycin. The transgenic tobacco plants could delay TMV infection for about 25 days compared with non-transgenic tobacco plants. Pokeweed antiviral protein Ⅱ is expressed with high level in summer leaves. The expression of PAPⅡ is regulated by season. The total RNA was extracted from pokeweed (Phytolacca americana L.) leaves in summer using the method of TRIzol and used as template to amplify the PAPⅡ gene by RT-PCR and then the gene was cloned into E. coli expression vector and secreted expression pPIC9K vector. The two vectors with PAPⅡ gene were then transferred into E. coli strain BL21 (DE3)-plysS and Pachia pastoris GS115 strain respectively. The specific protein was produced induced by IPTG and methanol.

由于美洲商陆抗病毒蛋白Ⅱ是一个受季节调控表达的蛋白，本实验以美洲商陆夏季叶片为材料，通过对其总RNA的提取、反转录、并用PAPⅡ的特异引物进行PCR扩增，对PAPⅡ进行了基因克隆、序列分析，结果扩增出来的PAPⅡ基因与已经报道的序列同源性是99.9％，然后将该基因构建到原核、真核表达载体上，分别转化了大肠杆菌和毕赤酵母并对它们进行了诱导表达，在两个表达系统中均获得了有活性的PAPⅡ表达蛋白，体外生物测定表明表达的蛋白均具有抑制病毒的活性，PAPⅡ基因在酵母中还没有表达的报道，这为获得具活性PAPⅡ蛋白提供了一种简便可行的方法。

The transgenic tobacco plants could delay TMV infection for about 25 days compared with non-transgenic tobacco plants.Pokeweed antiviral protein II is expressed with high level in summer leaves. The expression of PAPII is regulated by season. The total RNA was extracted from pokeweed (Phytolacca americana L.) leaves in summer using the method of TRIzol and used as template to amplify the PAPII gene by RT-PCR and then the gene was cloned into E.coli expression vector and secreted expression pPIC9K vector. The two vectors with PAPII gene were then transferred into E.coli strain BL21 (DE3)-plysS and Pachia pastor is GS115 strain respectively. The specific protein was produced induced by IPTG and methanol.

由于美洲商陆抗病毒蛋白Ⅱ是一个受季节调控表达的蛋白，本实验以美洲商陆夏季叶片为材料，通过对其总RNA的提取、反转录、并用PAPⅡ的特异引物进行PCR扩增，对PAPⅡ进行了基因克隆、序列分析，结果扩增出来的PAPⅡ基因与已经报道的序列同源性是99.9％，然后将该基因构建到原核、真核表达载体上，分别转化了大肠杆菌和毕赤酵母并对它们进行了诱导表达，在两个表达系统中均获得了有活性的PAPⅡ表达蛋白，体外生物测定表明表达的蛋白均具有抑制病毒的活性，PAPⅡ基因在酵母中还没有表达的报道，这为获得具活性PAPⅡ蛋白提供了一种简便可行的方法。

Result: Immunohistochemic results indicate that after HBO treatment, the expression of CytC and Bax in the neuronal cytoplasm decline significantly ( P.05) except in the time of 72 hours after TBI; the Bcl-2 expression is up-regulated significantly(p.05) after TBI. The images of electromicroscope indicate that before 24hr after TBI, mitochondria of control groups become more swoln even fracted and the matrixes become vaguer than those of being treated with HBO.

郑州大学2004年硕士研究生毕要论文高压氧对大民实脸性脑顶伤后神经细胞内cytochr她eC，Bol一2以及Bax表达的影响结果：(1)对于致伤后的脑组织切片染色发现，相同时间段内(3，6，12，24小时)实验组神经细胞内CytC阳性表达明显低于对照组(P.05)，而72小时以后无明显差异；Bel一2阳性表达于3，6，12，24，72小时时明显高于对照组(P.05)，Bax阳性表达于3，6，12，24小时时明显低于对照组，72小时时无明显差异。

①Rat MSC and VSMC were cultured and identified, respectively. MSC were labeled with DAPI firstly, and then co-cultivated with VSMC. The changes of morphology and ultrastructure of co-cultured cells were observed. Immunfluorescence analysis was performed by using monoclonal antibodies against specific antigen.②We established the regulatable system in two steps: a stable MSC line expressing rtTA has been constructed and characterized firstly by transfected with pUHD 17-1hyg and then selected by hygromycin B; in a second step, this line was used for trandfer the AT2R gene to MSC to get the well establishing double stable MSC lines;③The expression of AT2R regulated by doxycycline was evaluated by western blot;④The MSCs were transduced into rat carotid arteries with regulatable AT2R gene after the establishment of rat carotid balloon injury restenosis model. The intimal/medial area ratio were measured by digital analysis system.

研究方法：(1)密度梯度离心法及胶原酶消化法分别培养原代大鼠MSC及VSMC，细胞共培养并行免疫荧光化学染色和透射电镜观察超微结构；(2)组成受Dox调控的哺乳动物表达系统的四种成分的转化、扩增及提纯并酶切鉴定；(3)采用常规分子生物学方法连续两个回合转染体外培养的MSC，并分别采用发光计检测不同细胞克隆萤光素酶活性改变以及RT-PCR方法检测AT2R目的基因mRNA表达情况，根据各个细胞克隆受Dox调控表达的程度，选择低背景、高诱导表达AT2R的细胞系，作为双重稳定MSC细胞系；蛋白免疫印迹法观察该细胞系在Dox调控下AT2R表达的时相性、持续性及在不同浓度Dox调控下的表达情况；(4)建立大鼠颈动脉球囊损伤动物模型，将双重稳定MSC在术中导入血管，分别于14 d、28 d进行病理切片，检测可调控AT2R对新生内膜增生的影响；采用RT-PCR免疫组织化学免疫荧光等技术观察AT2R基因在新生内膜中的表达以及细胞外基质成分表达的改变，TUNEL法检测血管组织中细胞凋亡的变化情况。

In the present paper, the Sox gene expression analysis of different tissues from the Trionyx sinensis was studied by using RTPCR, and Sox gene fragments of expression from the testicle, brain and spleen were cloned using RTPCR products. The results show that Sox gene has specific expression in the testicle, brain, spleen, cardiac muscle and kindney and hasn't expression in muscle, liver and ovary. The results of clone reveal that the Sox genes of expression in the testicle are TSSox1 and TSSox 4, and those are TSSox2 and TSSox4 in the brain, and that is TSSox4 in the spleen. This suggests that the Sox gene act important role not only on the sex determination, but also on the development of neural system, immunocyte system and the differentiattion of male germ cell.

本文采用RT—PCR技术，研究了中华鳖不同组织Sox基因的表达，并通过PCR直接克隆法，分析了来自睾丸、脑和脾组织中的Sox基因序列结果表明在中华鳖的成体组织中，Sox基因在脑、心肌、肾、脾和雄性的睾丸组织中均有不同程度的表达，而在肌肉、肝脏和雌性的卵巢中则无表达，显示该基因具有组织表达特异性克隆分析显示，在睾丸组织中表达的是TSSox1和TSSox4基因，而在脑组织中表达的是TSSox2和TSSox4基因，脾组织中表达的是TSSox4基因此结果表明Sox基因不仅在性别决定中起作用，还可能在神经系统、免疫系统多种组织中起重要作用

The objectives of this study are:(1) To amplify Tb wbbL gene encoding rhamnosyl transferase bypolymerase chain reaction from the genomic DNA ofM.tuberculosis H37RV strain;(2) To clone PCR product of Tb wbbLgene into a cloning vector pMD18-T for sequencing;(3) To subcloneTb wbbL gene to an expression vector pET16b to construct pET16b-Tb wbbL; and to overexpress Tb WbbL protein in E.coli BL21(DE3)under different induction conditions;(4) To establish theco-expression system for expressing chaperons of pKJE7 plasmidand soluble Tb WbbL protein in E.coli BL21(DE3) under differentinduction conditions;(5) To test expressed WbbL protein by SDS-PAGEand Western blot methods.

本论文的目的是：(1)利用 PCR 方法从结核分枝杆菌 H37Rv 菌株的基因组DNA 中扩增出 Tb wbbL 基因；(2)将 Tb wbbL 基因克隆到pMD18-T 克隆载体中，经 DNA 序列测定证实为正确的基因；(3)将 Tb wbbL 基因亚克隆到 pET16b 表达载体中并通过改变不同的诱导条件在大肠杆菌 BL21(DE3)中表达 Tb WbbL 蛋白质；(4)建立在 BL21(DE3)大肠杆菌中共表达 Tb WbbL 蛋白质与分子伴侣的体系，优化表达条件以高效表达出可溶性 Tb WbbL蛋白质；(5)用 Western blot 方法鉴定所表达的 Tb WbbL 蛋白质为 Tb wbbL 基因产物。

In the aspect of research and development of HAS, the United States, Japan, united Kingdom, France and Italy recently tried tk use lower eucaryote, espesially yeast expression system to express HAS, obtained outstanding progress.

HSA的表达及工程化：将含HSA基因的酵母转化子单个菌落分别接种，并且分别进行甲醇诱导表达；将酵母表达的HSA蛋白质经SDS-PAGE电泳鉴定，筛选出多拷贝整合和高水平表达HSA菌株；将高表达HSA工程菌加保护剂后分别在液氮和－70C°低温冰箱内保存。

objective:to investigate the expression and significance of rb in the occurrence, development and regression of infantile hemangiomas.methods:the expression of rb was examined in the proliferative stage and catagen of human hemangiomas and normal skin tissues by using immunohistochemical technique.immunohistochemical technique for factorⅷ-related antigen was used to prove that the cells which expressed rb were endothelium.image analysis system was applied to measure the expression level of rb at different stages of hemangiomas and in normal skin tissues.results:the expression of rb was significantly lower in proliferating hemangiomas than that in involuting hemangomas(p.05).conclusion:rb might play an important role in the regression of human hemangioma endothelial cells and anti-angiogenesis.

目的：探讨rb蛋白在血管瘤发生、发展及退化过程中的表达状况及其意义。方法：采用免疫组织化学方法检测人皮肤血管瘤增生期、退化期及正常皮肤组织中rb的表达水平，并结合第ⅷ因子相关抗原的免疫组织化学染色证实表达rb的细胞是血管内皮细胞。利用计算机图像分析技术测量不同时期血管瘤组织和正常皮肤组织rb表达的积分光密度和面积。结果：增生期血管瘤内皮细胞rb表达水平低于退化期，差异有显著性（p.05），退化期血管瘤内皮细胞rb表达水平与正常皮肤组织相比，差异有显著性（p.05）。结论：rb通过抑制血管瘤内皮细胞增殖和血管生成而在血管瘤的退化过程中起重要作用。

Results: In cellular soluble protein electrophoresis,specific band about 75 KDa was observed in 593 strain.The differences of protein expression at 85,60,50,40KDa between 593 strain and 18 strain were observed.In extracellular soluble protein electrophoresis,specific bands at 75 KDa and 40 KDa were observed in 593 strain;specific band at 50 KDa was observed in 18 strain.The difference of protein expression amount in about 60KDa between 593 strain and 18 strain was observed.

结果：菌体可溶性蛋白电泳图中见593号菌株在75 KDa左右存在特异表达的蛋白条带，且二者在85、60、50、40 KDa左右蛋白表达量上存在差异；菌外可溶性蛋白电泳图中见593号菌株在75、40 KDa左右有特异表达的蛋白条带，而18号菌株在50 KDa左右有特异表达的蛋白条带，二者在60 KDa左右蛋白表达量上存在差异。

Neither the killing of Mr Zarqawi nor any breakthrough on the political front will stop the insurgency and the fratricidal murders in their tracks.

在对危险的南部地区访问时，他斥责什叶派民兵领导人对中央集权的挑衅行为。

In fact,I've got him on the satellite mobile right now.

实际上 我们已接通卫星可视电话了