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Interaction between fusion peptide of influenza virus A and its related antisense peptides was studied. A peptide was screened out with high binding ability to fusion peptide and the dissociation constant was shown at the level of umol/L. In order to quantitatively characterizing the affinity interaction, a system consisted of quartz crystal microbalance biosensor and flow injection analysis was designed and established.

据此初步结果,研究了甲型流感病毒融合肽与其反义肽的相互作用;提出了基于反义肽的组合化学法筛选目的多肽或蛋白质的亲和配基的全新的研究思路,筛选到了与融合肽具有强相互作用的多肽,其与融合肽的解离常数达到umol/L。

The shorter internodes and out-growth of anxillary buds found in antisense LeETR2 and leaf epinasty observed in antisense LeETRl plants are phenotypes consistent with the negative regulation model of ethylene.

5植株衰老和叶片脱落延迟,花瓣脱落也明显的延迟,在转反义LeETR1基因番茄的果实成熟特性被明显改变,表现在乙烯释放高峰延迟,果实颜色出现变异,以及和成熟相关的酶活性的改变,然而,转反义LeETR2基因的果实的成熟特性与对照相比无明显差别。

The stable clones are further identified by RT-PCR and Western blot; 6 MTT assay is used to investigate the effect of ZNRD1 on the cell growth of cells (AGS, SGC7901, MKN28, NIH3T3, GES-1); 7 Soft agar assay is used to investigate the effect of ZNRD1 on the clonality of cells (AGS, MKN28); 8 Nude mice assay is used to investigate the effect of ZNRD1 on the cell growth of gastric cancer cells (AGS, MKN28); 9 Flow cytometry is used to investigate the effect of ZNRD1 on the cell cycle distribution of cells (AGS, MKN28, NIH3T3, GES-1); 10 Flow cytometry is used to investigate the effect of ZNRD1 on the cell apoptosis of cells (AGS, MKN28, NIH3T3); 11 MTT assay is used to investigate the effect of ZNRD1 on the drug sensitivity of cancer cells (SGC7901, SGC7901/VCR, HL-60, HL-60/VCR) in vitro; 12 SRCA is used to investigate the effect of ZNRD1 on the drug sensitivity of gastric cancer cells (SGC7901, SGC7901/VCR) in vivo; 13 Flow cytometry is used to investigate the effect of ZNRD1 on adriamycin accumulation of cancer cells (SGC7901, SGC7901/VCR, HL-60, HL-60/VCR); 14 Transmission electron microscope is used to investigate the effect of ZNRD1 on the sensitivity of SGC7901 cells towards drug-induced apoptosis; 15 Flow cytometry and DNA ladder assay are used to investigate the effect of ZNRD1 on the sensitivity of cells (SGC7901, SGC7901/VCR, HL-60/VCR) towards drug-induced apoptosis; 16 Microarray is used to investigate the profiling of ZNRD1-responsive genes in gastric cancer cells (AGS, MKN28, SGC7901, SGC7901/VCR); 17 RT-PCR and Western blot are used to identify the results of microarray; 18 Reporter gene assay is used to investigate the effect of ZNRD1 on the transcriptional activity of cyclin D1; 19 Reporter gene assay is used to investigate the effect of ZNRD1 on the transcriptional activity of MDR1; 20 Kinase assay is used to investigate the effect of ZNRD1 on the activity of cyclin E-CDK2 kinase; 21 The antisensenucleic acids of p21 is used to inhibit the expression of p21, and flow cytometry is used to investigate the effect of p21 on ZNRD1-induced cell cycle arrest in gastric cancer cells; 22 The antisensenucleic acids of p27 is used to inhibit the expression of p27, and flow cytometry is used to investigate the effect of p27 on ZNRD1-induced cell cycle arrest in gastric cancer cells; 23 Liposome is used to up-regulate the expression of Skp2, and flow cytometry is used to investigate the effect of Skp2 on ZNRD1-induced cell cycle arrest in gastric cancer cells; 24 Western blot is used to investigate the effect of ZNRD1 on the stability of Skp2 and p27 in gastric cancer cells; 25 MVD assay is used to investigate the effect of ZNRD1 on the angiopoietic activity of gastric cancer cells; 26 ELISA is used to investigate the effect of ZNRD1 on the expression of VEGF165 in gastric cancer cells; 27 The roles of DARPP-32 in MDR of gastric cancer cells are investigated using gene transfection, MTT assay, SRCA, flow cytometry and DNA ladder assay.

应用杂交瘤技术制备ZNRD1的首个单克隆抗体;2)利用RT-PCR、Western blot和免疫组化检测ZNRD1在胃癌组织、胃炎组织、正常胃上皮组织、胃癌细胞和正常胃组织上皮细胞中的表达;3)构建ZNRD1的小干扰RNA载体,并测序鉴定;4)利用脂质体将ZNRD1的真核表达载体及其空载体转染胃癌细胞(AGS、SGC7901、MKN28)和小鼠成纤维细胞(NIH3T3),G418筛选后进行鉴定;5)利用脂质体将ZNRD1的小干扰RNA载体及其空载体转染药敏胃癌细胞(SGC7901)、正常胃组织上皮细胞(GES-1)、对长春新碱耐药的胃癌细胞(SGC7901/VCR)、药敏白血病细胞(HL-60)、对长春新碱耐药的白血病细胞(HL-60/VCR),G418筛选后进行鉴定;6)利用MTT实验检测ZNRD1高/低表达对细胞(AGS、SGC7901、MKN28、NIH3T3、GES-1)生长的影响;7)通过软琼脂克隆形成实验检测上调ZNRD1对AGS、MKN28细胞克隆形成能力的影响;8)通过裸鼠成瘤实验检测上调ZNRD1对AGS、MKN28细胞体内成瘤性的影响;9)通过流式细胞仪分析ZNRD1高/低表达对细胞(AGS、MKN28、NIH3T3、GES-1)的细胞周期的影响;10)通过流式细胞仪分析上调ZNRD1对细胞(AGS、MKN28、NIH3T3)的凋亡的影响;11)通过MTT实验检测ZNRD1高/低表达对细胞(SGC7901、SGC7901/VCR、HL-60、HL-60/VCR)体外药物敏感性的影响;12)通过肾包膜下移植法检测ZNRD1高/低表达对细胞(SGC7901、SGC7901/VCR)体内药物敏感性的影响;13)通过流式细胞仪分析ZNRD1高/低表达对细胞(SGC7901、SGC7901/VCR、HL-60、HL-60/VCR)内阿霉素蓄积和泵出的影响;14)通过透射电镜检测上调ZNRD1对SGC7901细胞凋亡敏感性的影响;15)通过流式细胞仪和DNA梯度试验检测ZNRD1高/低表达对细胞(SGC7901、SGC7901/VCR、HL-60)凋亡敏感性的影响;16)通过基因芯片检测ZNRD1高/低表达对胃癌细胞内基因表达谱的影响;17)利用RT-PCR、Western blot对基因芯片的结果进行鉴定;18)利用报告基因实验检测ZNRD1对cyclin D1的启动子活性的调节作用;19)利用报告基因实验检测ZNRD1高/低表达对MDR1的启动子活性的调节作用;20)利用激酶试验检测ZNRD1对cyclin E-CDK2 激酶活力的影响;21)利用反义核酸技术抑制p21的表达;通过流式细胞仪检测抑制p21对ZNRD1介导的细胞周期阻滞的影响;22)利用反义核酸技术抑制p27的表达;通过流式细胞仪检测抑制p27对ZNRD1介导的细胞周期阻滞的影响;23)利用脂质体转染法上调Skp2的表达;通过流式细胞仪检测上调Skp2对ZNRD1介导的细胞周期阻滞的影响;24)利用Western blot检测ZNRD1对p27和Skp2的蛋白稳定性的影响;25)利用微血管密度实验检测ZNRD1对AGS、MKN28细胞裸鼠移植瘤微血管形成的影响;26)利用ELISA检测ZNRD1对AGS、MKN28细胞培养上清和移植瘤匀浆中VEGF165含量的影响;27)利用脂质体转染法、MTT实验、肾包膜下移植法、流式细胞仪和DNA梯度试验检测新耐药相关分子DARPP-32对细胞(SGC7901、SGC7901/VCR、对阿霉素耐药的胃癌细胞SGC7901/ADR)多药耐药表型的影响;利用脂质体转染法和MTT实验检测下调ZNRD1对DARPP-32介导的胃癌多药耐药的调控作用。

However, the binding patterns between the HU-induced and uninduced HEL cells were different. The data in Southwestern-blot analysis showed that the molecular weights of the GATA factors binding to the HS2core DNA sequence were 43kd and 47kd respectively.

反义寡核苷酸技术及RT-PCR的结果表明,GATA-1的反义核苷酸抑制了羟基脲诱导的β-珠蛋白基因的表达,进一步证明了GATA-1因子在羟基脲诱导的HEL细胞分化过程中是不可缺少的促进因素。

Objective: To explore the mechanisms resulting in the recurrence of urethral scar which make urethral strictures difficult to be cured, a series experiments were conducted to find potential effective factors involved in urethral scar formation and degradation, including the studies of extracellular matrix component of urethral stricture scar, the characteristics of urethral scar fibroblast, and the effects of urine on urethral fibroblast in vitro, as well as the studies to compare the difference of collagenase activity, type Ⅰ collagen and tissue inhibitor of metalloproteinase-1 (TIMP-1) in the tissues and cultured fibroblasts from normal urethra and strictured urethra respectively, and the studies to investigate the effect of antisense TIMP-1 oligodeoxyonucleotide on cell proliferation and collagenase activity of urethral scar fibroblast.

中文题名尿道瘢痕基础研究副题名胶原酶活性,TIMP-1的表达及其反义基因治疗外文题名 Experimental study on urethral scar-activity of collagenase,expression of TIMP-1,and antisense TIMP-1 gene transfection of urethral scar fibroblast 论文作者黄翔导师杨宇如教授学科专业外科学研究领域\研究方向学位级别博士学位授予单位四川大学学位授予日期2002 论文页码总数104页关键词尿道手术瘢痕胶原酶成纤维细胞尿道瘢痕馆藏号BSLW /2003 /R699 /12 目的:研究尿道瘢痕的细胞外基质的组成,尿道瘢痕成纤维细胞的生物学特性以及尿液对其生长的影响;比较胶原酶活性,金属蛋白酶组织抑制因子-1(TIMP-1)以及Ⅰ型胶原含量在尿道瘢痕和正常尿道组织及体外培养的成纤维细胞中的差异;研究反义TIMP-1寡核苷酸对尿道瘢痕成纤维细胞增殖以及胶原酶活性的影响。

In this study,According to known sequence of Arabidopsis thalian cold-induced promoter rd29A,we cloned a 953bp distinctive fragement.

由于本研究中采用的是反义基因技术,个别碱基的突变并不影响该基因在反义RNA中的功能。

Tuberization of sense and antisense transgenic plants grown in vitro demonstrated that four of the ten sense plant lines had a significant increase while six of eleven antisense plant lines had a significant reduction in percentage of plantlets formed tubers and number of tubers per plantlet compared with the control.

对转正反义基因植株试管块茎形成的诱导结果表明,在10个转正义patatin基因植株中,有4个株系的结薯株率和单株结薯数比其对照明显增加,在11个转反义patatin基因植株中,有6个株系的结薯株率和单株结薯数比对照明显减少。

The mc2155 carrying pMind-glmU-AS was induced by different concentration of tetracycline, and GlmU protein was detected in the mc2155 carrying pMind-glmU-AS by Western blotting. The results showed that tetracycline of 20ng/ml has obviously inhibited the growth rate of mc2155 carrying pMind-glmU-AS, but the amount of GlmU protein did not change compared to that of uninduced mc2155 carrying pMind-glmU-AS.

glmU反义RNA的诱导表达与GlmU蛋白的检测分别用5 ng/ml、10 ng/ml和20 ng/ml的四环素诱导携带pMind-glmU-AS表达载体的mc~2155菌株表达反义RNA,绘制细菌生长曲线,结果表明20 ng/ml浓度的四环素可抑制mc~2155/pMind-glmU-AS的生长,但Western Blotting分析结果显示经四环素诱导和未诱导的mc~2155/pMind-glmU-AS中,GlmU蛋白表达量无明显的变化。

Used the method of touch down PCR, the ACS2 gene was cloned from DNA of Petunia hybrida. At the same time, its gene sequence DQ090003 was obtained in GenBank, the antisense repetition of ACS2 linked the plasmid pBI-121 by use of one step. In the result, the expression vector having antisense repetition gene was constucted.

利用降落PCR的方法,从矮牵牛DNA中扩增出ACS2基因片段,在GenBank中获得基因序列号为DQ090003;利用一步法将ACS2基因的反义重复与pBI-121质粒连接,构建含该基因的反义重复表达载体。

2The method of scientific investigation is nothing but the expression of the necessary mode of working of the human mind.

译文]:科学研究的方法只是人类思维活动的必要表达方式。注意:反义正译是重要的翻译方法之一。常见的反义正译的语言现象还有

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We have no common name for a mime of Sophron or Xenarchus and a Socratic Conversation; and we should still be without one even if the imitation in the two instances were in trimeters or elegiacs or some other kind of verse--though it is the way with people to tack on 'poet' to the name of a metre, and talk of elegiac-poets and epic-poets, thinking that they call them poets not by reason of the imitative nature of their work, but indiscriminately by reason of the metre they write in.

索夫农 、森那库斯和苏格拉底式的对话采用的模仿没有一个公共的名称;三音步诗、挽歌体或其他类型的诗的模仿也没有——人们把&诗人&这一名词和格律名称结合到一起,称之为挽歌体诗人或者史诗诗人,他们被称为诗人,似乎只是因为遵守格律写作,而非他们作品的模仿本质。

The relationship between communicative competence and grammar teaching should be that of the ends and the means.

交际能力和语法的关系应该是目标与途径的关系。

This is not paper type of business,it's people business,with such huge money involved.

这不是纸上谈兵式的交易,这是人与人的业务,而且涉及金额巨大。