细胞基因

Along with the fluconazole solution density rise,the experimental two kind of strain various glucose density is higher,showe d the glucose consumption are less,takes the logarithmof the medicine de nsity,discovered the logarithm the medicine density and each glucose den sity presents the linear relations;Carries on the analysis comparison to under the fluconazole function two kind of strain linear relations,disc overed the relations of the two strains has the nonuniformity.3 Compare the fluconazole induction reaiatance SC5314 strain and sens itive strain compares,its difference gene expression mainly concentrates in:The code proteinase body and the protein hydroltyic enzyme gene,in the code sugar fat metabolism process is connected the protein gene,the cell cycle correlation gene,the duplication and the translation adjustme nt correlation gene,the stress response correlation gene,the line plast ochondria correlation gene,the cell wall function related gene.4 Candida albicans SC5314 induction resiatance strain was processed b y Xianglian solution,its expression change gene mainly is:Code stress re sponse family protein gene,biomembrane relevant gene,a code proteinase body gene race,code cell cycle related protein gene,duplication and tra nslation adjustment related protein gene.5 The clinical reaiatance strain Candida albicans was processed by Xi anglian solution,its expression change gene mainly is:Codes the hot sho ck protein gene,the serine/threonine protein activating enzyme gene,the proteinase body family gene,the regulation copies and translates the ge ne.

随着氟康唑药液的浓度上升，试验的两种菌株各孔葡萄糖浓度越高，说明葡萄糖消耗越少，经过药物浓度取对数后进行分析，发现取对数后的药物浓度和每孔中葡萄糖浓度者呈现线性关系；对氟康唑作用下的两种菌株的线性关系进行分析比较，发现对两种菌株作用具有不一致性。3氟康唑诱导的耐药SC5314菌株与诱导前的敏感株相比，其差异基因表达主要集中在：编码蛋白酶体及蛋白水解酶的基因，编码糖脂代谢过程中相关蛋白的基因，细胞周期相关基因，转录及翻译调节相关基因，应激反应相关基因，线粒体相关基因，细胞壁功能相关基因。4白念珠菌SC5314诱导耐药株经香莲外洗液作用后，其表达变化的基因主要是：编码应激反应家族蛋白的基因，生物膜相关性基因，编码蛋白酶体基因一族，编码细胞周期相关蛋白基因，转录及翻译调节的相关蛋白基因。5白念珠菌临床耐药菌株经香莲外洗液作用后，其表达变化的基因主要是：编码热休克蛋白基因，丝氨酸/苏氨酸蛋白激酶基因，蛋白酶体家族基因，调控转录及翻译基因。

METHODS: Tum5 gene was amplified from plasmid pSPORT1Sfi by PCR technique and subcloned into the expression plasmid of lentiviral vector, pGCFU, to generate the lentiviral expression vector, pGCFUTum5. The correct Tum5 gene was confirmed by endoenzyme digestion and sequencing. Recombinant lentiviruses were produced by 293T cells following the cotransfection of pGCFU Tum5 and packaging plasmids-pHelper1.0and pHelper2.0. The resulting recombinant lentiviruses (GCFUTum5) which carried Tum5 and EGFPgene were then used to infect human umbilical vein endothelial cells.

采用PCR技术从含有tumstatin基因的质粒克隆模板 pSPORT1Sfi钓取Tum5基因，并将基因克隆到慢病毒载体表达质粒pGCFU中，构建慢病毒载体表达质粒pGCFUTum5，通过酶切、测序验证Tum5基因后，将pGCFUTum5质粒和包装质粒pHelper1.0，pHelper2.0共同转染人胚胎肾上皮细胞系293T细胞，获得携带Tum5基因和EGFP基因的重组慢病毒GCFUTum5，并转染靶细胞人脐静脉血管内皮细胞。

By retrovirus vector the foreign mdr1 gene could be efficiently transferred into bone marrow mononuclear cells of mouse in vitro, and it was confirmed that mdr1 gene was expressing stably and effectively in cells; in short terms there is no significant influence on the bone marrow MNC by transfection of mdr1 gene,but in the long run(in our study we had surveyed 2 months),there is a decline in the expression of apoptosis factor Bax in the MNC which transfected by mdr1 gene,in favour of the recovery of the receptor mouse;on the other hand it also poses a blance problem between multiplication and apoptosis in the process of receptor hematopoiesis after transplanting HSC which was transfected with mdr1 gene.

通过逆转录病毒载体可在体外将外源性mdr1基因转入小鼠骨髓单个核细胞中，转染的mdr1基因能整合到骨髓单个核细胞基因组中并有功能性表达；mdr1基因转入小鼠骨髓单个核细胞在移植后短期内对受体造血细胞没有显著影响，但从长期看来(本研究观察2个月)转染了mdr1基因的造血干细胞的凋亡因子Bax表达降低，更有利于受体鼠造血的恢复；另一个方面它也同时提出一个问题，即mdr1基因转染造血干细胞在回输受体后造血过程的增殖凋亡平衡。

Methods The NGF, BDNF, and NT3 genes of rats were cloned, the eukaryote expression vectors were established, the three kind of recombinant vectors were used to transfect astrocytes, the positive cloned cells were cultured dilatedly after G418 sifting; using supernates of culture liquid of astrocytes modified by gene to culture PC12 or TrkB-PC12, the expression and its level of gene target cells aimed genes were measured by Western blotting or immunohistochemical method.

克隆大鼠NGF、BDNF和NT3基因，构建真核表达载体；3种重组载体分别转染星形胶质细胞，G418筛选后获得阳性克隆细胞进行扩大培养；取基因修饰星形胶质细胞培养液上清培养PC12细胞或TrkB-PC12细胞；用Western blotting杂交或免疫组织化学方法检测基因靶细胞目的基因的表达及其水平。

The PSCA_3 fragment was selected for its superior expression level in eukaryotic cells.Then the sig-PSCA_3-Fc-GPI genetic fragment was cloned into pVAX1-neo-IRES-GM/B7 vector to construct the final immunological inhanced DNA vaccine pVAX1-PSCA_3-FcGB. Immunofluorescence and flow cytometry were used to confirm the expression of PSCA_3 fragment by transfected into Cos7 cell.Finally,the anti-tumor effect of pVAX1-PSCA_3-FcGB was tested in murine prostate cancer model generated by RM-1 cell line.The animal was immunized with pVAX1-PSCA_3-FcGB DNA vaccine by intramuscular injection plus electroporation,pVAX1 and pVAX1-PSCA_1-FcGB plasmid were used as control.The inhibitory effect of tumor was investigated by observion of forming time,volume and inhibition ratio of tumor.Results:DNA sequencing conformed that the heterological PSCA fusion antigen fragment which was synchronized by overlapping-extending-PCR,was consistent to design.Enzyme digestion analysis showed that the 1 to 4 copies heterological PSCA fusion antigen fragments were constructed successfully.

方法(1)检索GenBank，选择包含人主要T细胞抗原表位序列的人PSCA基因片段，应用异种化抗原设计技术，保留人T细胞抗原表位，设计异种化PSCA融合抗原片段；(2)根据核酸序列按中心模板法设计引物，应用重叠延伸PCR技术拼接合成异种化PSCA融合抗原片段基因，以PCR、限制性酶切和DNA序列测定法进行鉴定：(3)利用DNA限制性内切酶BssHⅡ和MluⅠ酶切后粘端互补的特点，采用同尾酶法构建1—4拷贝异种化PSCA融合抗原片段(PNCA_1-PSCA_4)，并将上述片段分别插入真核表达载体pCI-neo-Fc-GPI中，转染293T细胞，借助免疫荧光+流式细胞术考察插入片段表达效率，最终选定PSCA_3片段进行下一步研究；(4)将sig-PSCA_3-Fc-GPI基因片段自pCI-PSCA_3-Fc-GPI质粒上切下，插入pVAX1-neo-IRES—GM/B7载体中，构建免疫增效DNA疫苗pVAX1-PSCA_3-FcGB，并应用转染Cos7细胞+免疫荧光/流式细胞术方法鉴定其在真核细胞中的表达情况；(5)给8周龄雄性C57BL/6小鼠皮下种植RM-1细胞，制备小鼠前列腺癌模型，并采用股四头肌肌肉注射+电脉冲法(Electroporation，EP)接种DNA疫苗质粒pVAX1-PSCA_3-FcGB，同时接种pVAX1空载体质粒和pVAX1-PSCA_1-FcGB质粒作为对照，通过观察计算免疫动物的成瘤时间、肿瘤体积和抑瘤率，来评价该DNA疫苗在小鼠体内的抑瘤效果。

In addition, baculovirus are noninfectious to vertebrates, and their promoters have been shown to be inactive in mammalian cells. Futhermore, the expression level of some recombinant proteins is up to 1000 mg/L. Finally, recombinant proteins are soluble and antigenically, immunogenically and functionally similar to their authentic counterparts. In this article, we used the transfer vector pBlueBac of AcNPV (Autographa californica Nuclear Polyhedrosis Virus) to construct the recombinant baculovirus AcNPV-GM-CSF by in vivo recombinantion.

本文中，我们首先利用苜蓿夜蛾核型多角体病毒带β-Galactosidase 基因标记的非融合蛋白基因转移载体pBlueBac，通过细胞内同源重组的方式将hGM-CSF基因成功地插入病毒AcNPV的基因组中。hGM-CSF基因编码信号肽和完整的氨基酸序列。hGM-CSF基因在感染重组病毒的草地夜蛾培养细胞Sf9中得到表达，感染后的Sf9细胞培养液能刺激人骨髓细胞在体外形成典型的集落，表达水平可达2.7〓CFU／ml。

Moreover, CoI gene was more conservative and a lower evolutionary rate than Cyt b gene. CoI gene was an effective marker in analysing the phylogenesis of families in Passeriformes. It can be used in identifying the species of Passeriformes, but it was revealed that CoI gene was more suitable to identify the phylogenetic relationship of avian family unit than Cyt b gene, and CoI gene can be a molecular marker to identify avian species. But in species identification, CoI gene was less stable and accurate than Cyt b gene. We suggested youd better employ the other marker if you have the CoI gene only.(2) In the phylogenetic trees of birds from Lanius, L.

比较分析了雀形目6科15种鸟类的细胞色素b全序列和CoI基因部分序列，结果显示细胞色素b和CoI基因序列的变异位点分别为454个和366个、简约信息位点为337个和303个，而且线粒体CoI基因比细胞色素b基因略微保守、进化速率也较低；CoI基因在确定雀形目科级阶元之间的系统发生关系方面是一种有效的分子标记，同时它也能够用于雀形目鸟类的物种鉴定，但在物种鉴别方面不如细胞色素b基因稳定、准确。

Methods At first, retrovirus vectors encoding IFN-γ or IL-4 gene were constructed. They were transfected into PA317 packaging cells by lipofectamine, PA317IFN-γ and PA317IL-4 cells were obtained. C6 glioma cells were infected with replication deficiency retrovius containing IFN-γ or IL-4 gene, cell morphology、cell growth curve and cloning efficiency assay were examined. The intracranial C6 glioma animal model was established in immunocompetent Wistar rats. PA317IFN-γ and PA317IL-4 cells were stereotactically implanted into the tumor areas alone or together. The survival of tumor bearing rats were examined, tumor volumes were measured and immunohistochemical analyses of CD4〓 and CD8〓 T lymphocyte infiltration were performed.

构建和鉴定携带IL-4或IFN-γ基因的逆转录病毒载体，将其导入逆转录病毒包装细胞PA317，通过G418抗性筛选，得到携带目的基因的包装细胞并鉴定；应用携带IL-4或IFN-γ基因的复制缺陷型逆转录病毒感染C6胶质瘤细胞，观察对C6胶质瘤细胞形态、细胞生长曲线和克隆形成率的影响；建立C6胶质瘤大鼠脑内移植动物模型，将携带IL-4和IFN-γ基因的逆转录病毒包装细胞分别或联合注射到脑内荷瘤大鼠的肿瘤组织中，观察其治疗作用，并初步探讨其机制。

In summary, the BRD7 gene acted as a candidate of tumor suppressor gene with NPC. The overexpression of BRD7 can partly reverse malignant phenotype of NPC cell line. The suppression effect of BRD7 on NPC tumorigenesis my be achieved by recognizing acetylated histone peptide through their motif-bromodomain, then modulating gene transcription by taking part in histone acetylating and chromosome remodeling, finally influencing signal-transduction pathways.

综上所述，BRD7基因作为一个重要的鼻咽癌抑瘤基因侯选者，在鼻咽癌细胞中的过表达后，可导致鼻咽癌细胞 HNE蛋白质表达谱发生改变，逆转其恶性表型，其作用机理可能是：BRD7基因通过其功能域彭聪硕士学位论文10Bromodomain与乙酚化的组蛋白特异性结合，参于染色体的乙酚化，染色质的组装，从而影响基因转录的调控，最终影响细胞内的信号传导通路并实现对细胞周期的调控，从而发挥抑制鼻咽癌细胞生长的作用。

At the same time, the process of the wound healing was observed histologically, and the regeneration cycle of epidermal cells and the temporal change in inflammatory cells were measured. Results Inflammatory cells infiltrated into wound surface were neutrophils, followed by macrophages and lastly lymphocytes. Epidermal cells proliferated most actively on PBD 3 and the mitoses of them increased significantly on day 7 after burn. The gene expression of PDGF, PDGFR and EGFR reached peaks on PBD 1 and the gene expression of EGF and TGFβ-R2 were highest on PBD 3. In addition, the gene expression of TGFβ-R1 and TGFβ1 increased significantly on PBDs 5 and 7 respectively.

结果 炎性细胞到达创面依次为中性粒细胞、巨噬细胞和淋巴细胞；表皮细胞周期变化为伤后3天细胞增殖活跃，伤后7天细胞分裂明显增多；伤后1，3天PDGF、PDGFR(血小板源性生长因子受体)和EGFR基因表达明显增强，伤后3，5天EGF、TGFβ-R2基因表达增强，伤后5，7天TGFβ-R1(转化生长因子β受体1)基因表达增强，伤后7，10天TGFβ1基因表达增强。

What are your goals and strategies for growth?

你的成长目标和策略是什么？

And unto the angel of the church in Sardis write; These things saith he that hath the seven irits of God, and the seven star I know thy works, that thou hast a name that thou livest, and art dead.

3：1 你要写信给撒狄教会的使者，说，那有神的七灵和七星的，说，我知道你的行为，按名你是活的，其实是死的。