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Methods Eukaryotic expression plasmid pCMVCD was constructed , and identified by re2 striction endoenzyme digestion. CD gene was transfected into NSCs from new2born Wistar rats using Lipofec2tamine2000. Positive clones (named NSCs/ CD cells) were screened by G418 presence. 52Fluorocytosine (52FC) ad2ministration of different concentrations were incubated with NSCs/ CD cells. NSCs/ CD cells viability ratios were mea2 sured by MTT assay.

通过构建真核表达质粒pCMVCD ,限制性内切酶消化鉴定后,采用Lipofectamine 2000 脂质体介导法转染新生大鼠室管膜下区神经干细胞(Neural stemcells , NSCs),G418 筛选阳性克隆,加入不同浓度的52氟胞嘧啶(52Flourocytosine , 52FC),MTT比色法测定NSCs的生存率。

Methods The BHD vector and the small interfer RNA targeting BHD was constructed and transfected into the cell line A549. The expression of BHD was examined by immunofluorescence. Transwell test was used to detect the invasion ability of A549 cells. Results After transfected BHD vector into A549 cells, immunofluorescence showed the expression of Folliculin located in cellular plasma.

制备BHD真核表达质粒,转染肺腺癌细胞,用免疫荧光法检测Folliculin蛋白在肺癌细胞中的表达位置;筛选sRNAi片段,选取有效抑制片段转入肺癌细胞,与转入BHD质粒的肺癌细胞对比,用Transwell实验了解BHD基因对肺癌细胞运动方面的影响。

Result: 1.(1) Three strips come to appear in the position of 6.0kb, 5.4kb and 600bp after gelose electrophoresis and their size accord with recombined plasmid DNA, pcDNA3 plasmid and VEGF165 gene accordingly.(2) Pure degree of the pcDNA3-VEGF165 recombined plasmid is: OD260/OD280=1.87. Its

在进行了重组质粒的扩增、提取、纯化、鉴定、兔MSCs的原代及传代培养以及RT-PCR、Western Blot 转染检测后,实验取得了令人满意的结果,证实了MSCs 能在体外有效转录外源VEGF 基因并分泌出VEGF 蛋白,同时也说明pcDNA3-VEGF165重组真核表达质粒是有效的。

Results Activities of SCs in the oxidative injured group decreased compared with the control group. However, these changes were significantly reversed by the IGF-1 pre-conditioning.

结果 与正常组及保护组相比,H2O2处理组细胞具有典型的凋亡形态特征:核染色质浓染、边集,细胞体积缩小,细胞存活率降低;SOD含量明显减少(P<0.01,MDA含量明显增加(P<0.01,Bcl-2表达下调;而IGF-1保护组细胞存活率明显升高(P<0.01,SOD含量较损伤组增高(P<0.01,MDA含量较损伤组明显减少(P<0.01,Bcl-2表达明显上调。

The hEGF was detectable in the supernatant of the modified keratinocytes. CONCLUSION The human keratinocytes modified with exogenetic hEGF cDNA can express and secrete hEGF.

由真核表达质粒pcDNA3-hEGF转染人角朊细胞获得的含外源性hEGF基因的角朊细胞具有表达分泌 hEGF的功能。

Objective:To recombine the plasmid PcDNA3.1-sTNFRⅠ,and detect the expression of recombinant plasmid in pericementum cell,and evaluate the bioactivity of expressed sTNFRⅠ.

目的:构建真核表达质粒PcDNA3.1-sTNFRⅠ,并转染人牙周膜细胞观察是否有目的产物的表达,研究其是否具有生物学活性,为进一步研究其在牙周治疗中的作用提供实验基础。

Full length LMP2A cDNA was firstly incised from pGEM-T-LMP2A with EcoR Ⅰ, Sma Ⅰ digestion, and then inserted into eucaryote. expression plasmid pCIcc controlled by CMV promoter. The CMV-LMP2A-SV40 expression unit was digested by ClaI, and inserted into E1-substituted adenovirus vector pAx1cw. Then the LMP2A recombinant adenovirus vector was cotransfected into 293 cells together with EcoT22I digested Ad5-TPC.

将带有LMP2A cDNA的重组质粒pGEM-T-LMP2A用EcoR Ⅰ、Sma Ⅰ双酶切下LMP2A cDNA,并将其插入含同样酶切位点的真核表达质粒pCIcc中,使其受控于CMV启动子下;用ClaI切下CMV-LMP2A-SV40表达单元,插入E1、E3区替代的腺病毒载体pAX1CW,选择正确的克隆pAX1CW-LMP2A与Ad5 DNA-末端肽复合体共转染293细胞,通过同源重组获得复制缺陷型的重组腺病毒(Ad5-LMP2A)。

RESULTS: All the four recombinant eucaryotic plasmids were constructed successfully, and HBV core protein expression was confirmed by internal reference for transfection.

结果: 4份重组真核表达质粒均构建成功,转染至HepG2后通过内参照EGFP可鉴定出均有HBV-C蛋白表达。

Objective To construct recombination eukaryon expression plasmid for human parathyroid hormone gene, assay PTH expression and biological activity after transfection in vitro and evaluate gene therapy effect on hypoparathyroidsm.

目的构建甲状旁腺激素基因的重组真核表达质粒,评价体外转染后PTH基因的表达与生物学活性,同时观察其对甲状旁腺功能减退动物的基因治疗作用。

OBJECTIVE: To transfect rabbit bone marrow mesenchymal stem cells using pIRES2-EGFP-BMP-2 eukaryotic expression plasmid.

目的:使用前期已经构建成功的pIRES2-EGFP-BMP-2真核表达质粒转染兔骨髓间充质干细胞。

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