查询词典 combinant

Then the combinant plasmid was conducted into ACC-2 cell by electroporation. ACC-2 cells stably expressing CD was obtained by 10-day positive selection with 400 μg/mL G418. Total RNA was extracted and the expression of the CD gene in transfected ACC-2 cells was identified by RT-PCR. RESULTS: PCR yielded a fragment of l280bp and CD was verified by sequence analysis.

测序正确后，将其亚克隆到质粒表达载体pIRES中，构建以内部核糖体进入位点相连的CD基因的质粒表达载体pIRES-CD，采用电穿孔法，以质粒表达载体转染ACC-2细胞，用400μg/mL的G418筛选10d，获得稳定表达CD基因的ACC-2细胞系，提取该细胞的总RNA，用RT-PCR检测CD基因的表达。

Methods The yield of combinant human IL-18 expression in E.coli remains quite low because the sequence of mature hIL-18 has 37aa rare condons for E.coli in a total of 157aa.

它们的分子中没有N-糖基化位点，不存在二硫键，很可能巯基就是分子的活性中心。

Objective: To contructre combinant plasmid pET28a/hES and coexpress human Endostatin and predhPK-5 in E. coli. Methods: mRNA from human liver tissue was exracted, and the Endostatin gene was amplified by RT-PCR.

目的构建人内皮抑素原核表达质粒pET28a/hES，并与简化人纤溶酶原饼环区(predigested human PlasminogenKringle5，predhPK-5)在大肠杆菌中实现共表达。

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。

Methods The yield of combinant human IL-18 expression in E.coli remains quite low because the sequence of mature hIL-18 has 37aa rare condons for E.coli in a total of 157aa.

设计特异性引物，以RT-PCR从外周血单个核细胞中扩增出人白细胞介素18成熟肽片段，克隆进入PBV220载体。

Methods:The total RNA was extracted from mouse RAW264.7 cells stimulated by LPS.The sequence including the whole length of HMGB1 was amplified by RT-PCR and inserted into pMD-19T.The combinant vector was used as a template for PCR which was cloned into vector pMD-19T,then subcloned into expression vector pET-26b with pelB signal sequence and His-Taq sequence.After transforming E.coli BL21(DE3) and four hours induction by IPTG,HMGB1 expression confirmed by SDS-PAGE and the purification was performed by Ni2+-chelate affinity chromatograph.

脂多糖刺激后的RAW264.7细胞，提取总RNA，经RT-PCR扩增出含HMGB1的目的片段，克隆于pMD-19T载体，再亚克隆至含有pelB引导肽及His-标签肽的高效表达载体pET-26b，转化大肠杆菌BL21（DE3），经IPTG诱导后行SDS-PAGE鉴定目标蛋白表达，用镍鳌合琼脂糖凝胶亲和层析法分离纯化含His-标签肽的目的蛋白。

Methods:The total RNA was extracted from mouse RAW264.7 cells stimulated by LPS.The sequence including the whole length of HMGB1 was amplified by RT-PCR and inserted into pMD-19T.The combinant vector was used as a template for PCR which was cloned into vector pMD-19T,then subcloned into expression vector pET-26b with pelB signal sequence and His-Taq sequence.

脂多糖刺激后的RAW264.7细胞，提取总RNA，经RT-PCR扩增出含HMGB1的目的片段，克隆于pMD-19T载体，再亚克隆至含有pelB引导肽及His-标签肽的高效表达载体pET-26b，转化大肠杆菌BL21（DE3），经IPTG诱导后行SDS-PAGE鉴定目标蛋白表达，用镍鳌合琼脂糖凝胶亲和层析法分离纯化含His-标签肽的目的蛋白。

The combinant vector was used as a template for PCR which was cloned into vector pMD-19T,then subcloned into expression vector pET-26b with pelB signal sequence and His-Taq sequence.After transforming E.coli BL21(DE3) and four hours induction by IPTG,HMGB1 expression confirmed by SDS-PAGE and the purification was performed by Ni2 -chelate affinity chromatograph.

脂多糖刺激后的RAW264.7细胞，提取总RNA，经RT-PCR扩增出含HMGB1的目的片段，克隆于pMD-19T载体，再亚克隆至含有pelB引导肽及His-标签肽的高效表达载体pET-26b，转化大肠杆菌BL21（DE3），经IPTG诱导后行SDS-PAGE鉴定目标蛋白表达，用镍鳌合琼脂糖凝胶亲和层析法分离纯化含His-标签肽的目的蛋白。

The GPA1 gene was obtained via PCR amplification and was cloned in the two-hybrid DNA binding domain vector pGBKT7 The combinant plasmid was designated as pGBKT7-Ga.β-galactosidase assay indicatedthat Got did not have the property of self-activation. After pGBKT7-Ga was transformed to yeast pJ69-4A, we transformed Arabidopsis vegetative tissue two-hybrid cDNA library plasmids to yeast pJ69-4A containing pGBKT7-Ga.

通过PCR扩增得到GPA1基因并将其克隆到双杂交DNA结合域载体pGBKT7中，得到的重组质粒命名为pGBKT7-Gα，β—半乳糖苷酶活性鉴定表明Gα不具有自激活特性，将其转化到酵母菌pJ69-4A中，再以此为受体菌转化拟南芥绿色营养组织cDNA文库质粒。

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。

The studies on the developing of new media are very important to the increasing of the area density of HDD and to the implementation of hybrid recoding technology.

光磁混合记录方法，是一种可以突破超顺磁极限的限制，并进一步提高硬盘记录密度和读写速率的一种新型超高密度信息存储方式。

In io, mTOR inhibition promotes T cell anergy under conditions that would normally induce priming.

体内研究中，在正常可激发免疫反应的条件下，抑制mTOR可促使T细胞无能。