重组

In order to evaluate the potential of this immunodominant peptide antigen in serodiagnosis of TB, we have undertaken cloning and expression of a large number of esat-6p1 genes in Escherichia coli and purification products.

本实验研究目的是通过构建结核菌ESAT-6蛋白免疫优势段P1的重组多表达载体，在大肠杆菌 BL21（DE3）中进行诱导表达，并对表达产物进行纯化和复性，以获得大量ESAT-6P1抗原，同时对重组多免疫反应性进行鉴定评价，为探索重组多在结核病血清学诊断的应用奠定前期实验基础。

Tilter of the recombinant vaccinia virus is 1×109 pfu/ml.Conclusion This experiment provides a new approach to study the function of CD20 which will also lay a foundation of preparation for the mAb against CD20. human CD20;recombinant vaccinia virus;gene expression

本实验将编码人CD20的cDNA重组到痘苗病毒载体pJSA1175中，构建含人CD20编码cDNA的重组痘苗病毒，希望获得能够表达CD20的重组病毒，用于CD20分子的功能研究及单抗或多抗研制。

Reverse Transcription-Polymerase Chain Reaction was used to clone the whole-length gene of BMP-2 and construct AdV vector.

应用反转录-聚合酶链反应法克隆出骨形态发生蛋白质类2全长基因，构建重组腺病毒载体，DNA-磷酸钙共沉法将辅助质粒PJM17转染293细胞，同源重组构建出重组腺病毒。

Recombinant adenovirus was identified by polymerase chain reColon cancer cell line SW480 was infected with recombinant adenovirus Ad-p27mt, and expression of p27 protein was detected by Western blot.of expressed p27 protein after Ad-p27mt transfected colon cancer cell.novirus framework plasmid pAdeasy-1 with pShuttle-CMV-hp27mt, 30％combinant adenovirus DNA contained the target gene.

Ad-p27mt的聚合酶链反应检测Ad-p27mt转染大肠癌细胞后的p27mt的表达。结果：①用含pShuttle-CMV-hp27mt转化含pAdeasy-1的超感受态BJ5183后，可获得约30％的阳性重组质粒。②聚合酶链反应检测表明重组腺病毒DNA中含有目的基因。重组腺病毒滴度为7.95×1015pfu/L。

The study showed the recombinant wt/mCREG protein depressed the VSMC proliferation depending on dose and the optimal concentration was 400nM;2biologic function of CREG protein and the membrance receptor mechanism:①effect on VSMC migration: the wound healing experiment showed the OB2 cells migration was slower significantly after added wt/mCREG(400Nm) in supernatant. The HITASY cells migration were very slowly and no remarkable change. The gelatinase digestion and Western blot analysis showed the matrix metalloproteinase was decreased and TIMPs was increased;②effect on differentiation: after added wt/mCREG(400nM), the expression of myocardin, SMα-actin, MHC and caldesmin were increased and that of LM-1 and FN were decreased in OB2 cells. These effects were more significant when adding wtCREG.;③effect on VSMC proliferation: Cell cycle assay and BrDU stain showed: after added the wtCREG and mCREG protein, the ratio of cell in G0/G1 phase increased to 0.5773 and 0.5572 from 0.5308 respectively in OB2 group, which increased to 0.7369 and 0.7034 respectively from 0.6297 in HITASY group;3Role of M6P/IGF2R in CREG biologic function:①ELISA and co-immunoprecipitation showed the wt/mCREG binding to M6P/IGF2R directly.②antibody blocking test: when the anti-IGF2R was added to medium at the same time with wt/mCREG at different concentration(2μg/mL、4μg/mL、8μg/mL),the effects of CREG protein which depressing proliferation, migration, secretion and promoting differentiation were blocked, which had the positive correlation to the concentration of added anti body. The studies showed two combinant CREG promoted VSMC switch to differentiation phaenotype, at the same time, depress VSMC proliferation, migration and secreting extracellular matrix.

上述实验结果证实：两种重组CREG蛋白对VSMC增殖均有剂量依赖性的抑制作用，并且相同浓度的糖基化的CREG蛋白对细胞增殖的抑制效应更为显著，最佳效应浓度为400nM；2两种重组CREG蛋白添加后对HITASY和OB2细胞生物学行为的影响：①CREG蛋白对VSMC迁移的影响：刮伤实验发现，加入最佳效应浓度的wtCREG和mCREG蛋白24h后，OB2组迁移能力下降，HITASY组无明显变化；细胞外基质金属蛋白酶-2，9(Matrix metallo-proteinase 2，9，MMP2 ，9)明胶酶电泳检测和Western blot检测结果证实，两种CREG蛋白均可以使OB2细胞合成细胞外基质MMP2，9减少，而组织金属蛋白酶抑制物(Tissue Inhibitors of Metalloproteinases，TIMPs)增加；②CREG蛋白对VSMC分化的影响：加入400nM的wtCREG和mCREG蛋白12h后，OB2细胞myocardin、SMα-actin、MHC、caldesmin表达增加，LM-1、FN表达减少；③流式细胞仪分析细胞周期和BrDU染色分析证实，加入400nM的wtCREG和mCREG蛋白后，OB2组G0/G1期细胞由0.5308分别增加至0.5773和0.5572，HITASY组G0/G1期细胞由0.6297分别增加至0.7369和0.7034；3M6P/IGF2R在重组CREG蛋白的生物学功能中的调控作用：①免疫共沉淀和免疫荧光双染色分析结果显示，CREG蛋白与M6P/IGF2R存在直接结合；②应用抗体阻断实验：将不同浓度的anti- M6P/IGF2R(2、4、8μg /mL)与两种CREG蛋白同时加入培养液中，CREG蛋白抑制VSMC增值、迁移和合成细胞外基质、促进分化的效应减弱，而且与加入anti- M6P/IGF2R浓度正相关。

Firstly, the recombinant engineering technology related vaccine preparation was set up, including: 1. the gene knock-out based on linear DNA target molecular in vivo; 2. the selection and counterselection technology of exogenous gene knock in definite site on chromosome; 3. the Gap-repair clone technology in vivo; 4. the recombination technology of single-chain DNA and the overlapping primer; 5. Set up the new selection and counterselection technology of neo-E.

本研究首先建立与疫苗制备相关的重组工程技术包括：①基于线型DNA打靶分子的大肠杆菌体内基因剔除技术，②选择与反选择外源基因定位敲入染色体上特定位点技术，③Gap-repair体内克隆技术，④单链DNA重组技术和重叠引物介导的DNA重组技术，⑤建立&抗生素-酶切位点&选择反选择技术新技术。

According to the classical signal transduction pathway of estrogen, we use Luciferase reporter gene to screen phytoestrogen form Selaginella tamariscina, Pinus massoniana Lamb, Corallodiscus flabellate, Dryopteris sublaeta Ching et Hsu and Leonurus heterophyllus. In this study,the pβgal-Control vector, the pERE-TAL-luc vector, the pCNX2-hERα/pCNX2-hERβvector were transfected into HEK293 cell line. If the components in the herbs combine with the ER, then it could induce the expression of reporter gene controlled by ERE.

根据体内的雌激素信号转导途径，我们选用了以luc为报告基因、ERE为调控序列的重组报告基因载体pERE-TAL-luc，并将其与重组人ERα(humanERα，hERα)表达载体pCXN2-hERα和重组人ERβ(humanERβ，hERβ)表达载体pCXN2-hERβ共转染ER阴性细胞系，建立了靶向ERα或ERβ的细胞水平药物筛选模型，结合小鼠子宫增重实验，对卷柏、松针、益母草、石胆草及贯众等5种中药的植物雌激素作用进行了研究，以期为临床防治更年期综合症等提供准确、科学的实验基础。

This work might lead to develop many better methods or means of preventing and controlling dysenteric diarrhea.To elucidate the role of H-NS protein in bacterial physiology, the hns gene insertion and deletion mutants were constructed with RecA and Red recombination system. Then, growth curve and invasion ability of wild strain and deletion mutant were studied.

为了更深入的了解hns基因在细菌生命活动中所起的作用，我们首先利用RecA重组系统构建了痢疾杆菌2457T hns基因的插入突变体；随后，对基于Red重组系统的痢疾杆菌2457T缺失突变体的构建方法进行了摸索并成功地敲除了hns、hdeA、hdeB、yhiE和yhiF 5个基因，研究表明延长同源臂的长度可以明显提高同源重组的效率，缩短构建缺失突变体所需的时间。

Then plant expression vector pBG128 contains a germin gene ,a Nptll gene as the selectable marker and a B -gluceronidase gene as the reporter gene.

但反向克隆的重组子得到的小片段大小为1，567bp，正向克隆的重组子所得的小片段大小为968bp，并命名正向克隆的重组子为pBG128。

The new guidelines for debt restructuring greatly standardized the behavior of corporate debt restructuring to prevent the reorganization of the enterprise through the machine to manipulate profits, glossing over the accounting statements, but still need to continue to consider.

新的债务重组准则极大的规范了企业的债务重组行为，防止了企业借重组之机操纵利润、粉饰会计报表，但仍然存在需要继续考虑的问题。

Means the parent directory, so this command means to execute "toolchain.sh," which is in the current directory.

代表父目录，所以这个命令就是执行当前目录下的"toolchain.sh"。

Yes,In fact,I'm on our city ream.

是的。事实上我是我们市队的。