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Two 15nt complementary ssDNA primers were ligated into double stranded satRNA.Complete sequence of satRNA was obtained by RT-PCR using these 15nt ssDNA as amplification primers.Phylogenetic analysis showed that the identity of this 383nt satellite and some documented satRNAs was 72.6 to 99.5% at the nucleotide level.

以常规引物对感病植物的总RNA进行RT-PCR扩增,获得CMV RNA3全长克隆,经测序显示该RNA3序列属于CMV亚组Ⅱ;通过在卫星dsRNA两端分别加上15nt的单链DNA接头,以接头互补序列为引物进行扩增获得一个383nt的卫星RNA的全长序列。

Cylinders with a full-length eductor tube, or what is sometimes called a full-length dip tube, have a tube that runs from the inlet of the cylinder valve to the bottom of the cylinder .

带有全长排放管,或者有时称为全长汲取管的钢瓶有一个从钢瓶入口一直伸到钢底部的管子。

The venom of elapid snakes contains a number of small proteins that display a broad spectrum of pharmacological activities.In study on the neurotoxin from the venom gland of Bungarus multicinctus,five cDNA sequences (MNCTL1-a,MNCTL1-b,MNCTL1-c,MNCTL1-d and MNCTL2) encoding two novel proteins were obtained from the total RNA by reverse transcription-polymerase chain reaction.

采用SMART技术和RT-PCR方法,从同一个银环蛇毒腺中克隆到了5 种尚未报道过的心脏毒素类似物全长cDNA序列(MNCTL1-a、MNCTL1-b、MNCTL1-c、MNCTL1-d和MNCTL2)。5种序列中,MNCTL1-b、MNCTL1-c、MNCTL1-d和MNCTL2的长度均为505 bp,包括5′非翻译区32 bp,3′非翻译区212 bp和其余261 bp组成的一个完整开放阅读框;MNCTL1-a 由于3′非翻译区24 bp的缺失,全长cDNA仅为481 bp。

Results (1) The forward and reverse subtracted cDNA libraries of different metastastic potential large cell lung cancer cell lines were successfully constructed;(2) With blue-white colony and dot blot, 307 positive clones in the forward subtracted library and 78 positive clones in the reverse subtracted library were obtained;(3) 55 clones were successfully sequenced in the forward subtracted library. Homolog analysis confirmed 23 differential expression segments, which were similar to the human genes already known, including NME2, NPM1, MT 2A, HSPE1, TAFIA, EPRS, PX19 and EIF3S9 et al;(4) 31 clones were successfully sequenced in the reverse subtracted library. Homolog analysis confirmed 16 differentially expressed segments. 15 of them were similar to the human genes already known, including ANXA2, TUBB, PKN2, GNAS, EEF1A1, SSR2 and RPLPO et al. Only one segment had partial homology to known human genes. This segment was supposed to be the new EST segments which might not have been cloned.

结果(1)成功构建人大细胞肺癌高低转移株差异表达基因正向消减cDNA文库和反向消减cDNA文库;(2)经蓝白菌落筛选和斑点杂交,正向消减文库获得307个阳性克隆,反向消减文库获得78个阳性克隆;(3)正向消减文库挑选55个克隆成功测序,经同源性分析最终确定差异表达基因片段23个与已知人类全长基因具有很高相似性(95%~100%),这些基因包括NME2、NPM1、MT2A、HSPE1、TAFlA、EPRS、PX19和EIF3S9等;(4)反向消减文库挑选31个克隆成功测序,经同源性检索和比对最终确定差异表达基因片段16个,其中15个与人类全长基因具有很高相似性(95%~100%),包括ANXA2、TUBB、PKN2、GNAS、EEF1A1、SSR2和RPLPO等基因。1个片段和己知人类基因仅有部分相似性,表明它可能是未被克隆的人类基因EST片段。

Six different deletion and insertion mutants of BBSV-X were constructed and these were inoculated to Chenopodium amaranticolor. The preliminary results show that the sat-RNA could enhance symptom of BBSV-X and its insertion mutants on Chenopodium amaranticolor, but the pathogenicity of the CP deletion mutants was reduced.

构建了BBSV-X基因组6个缺失和插入突变体,将它们的体外转录物单独或与sat-RNA一起混合接种苋色藜,初步实验结果表明,sat-RNA能增强全长BBSV-X基因组及含全长基因组的突变体对苋色藜所致的症状,但减轻CP缺失突变体的致病性。

Food compositions of M. albus were investigated in areas of three counties in Hubei province. Diets of the M. albus with body length less than 100mm change with their growth: from Rotifera and Cladocera to Oligochaeta and Chironomidae.

通过对不同产地的271尾野生黄鳝的食性的研究表明,全长小于100mm的稚鱼,其食性随着全长的生长而变化,稚鱼前期以摄食轮虫、枝角类为主,后期以水生寡毛类、摇蚊幼虫为主。

Full-length cDNA of Gibel carp pou2 gene was cloned from gastrula SMART cDNA library through RACE-PCR. It consists of 2421 bp, with an ORF of 1416 hp encoding a protein of 471 aa. The complete amino acid sequence shares 91.0% identity with zebrafish pou2 gene product.

用RACE-PCR方法从原肠期SMART文库中扩增到银鲫pou2基因的全长cDNA,其全长为2421bp,开放阅读框为1416bp,编码471个氨基酸,与斑马鱼pou2基因的氨基酸序列一致性高达91.0%。

In the research, one kind of guzmania species named ostara was employed to be as material, and the full-length cDNA plasmid library of floral organ was constructed successfully by using SMART technology. The library had 3×10^6 original titer and more than 1 kb insert fragments. After 5'EST sequencing from 2004 positive clone chosen at random and clustering analyse, 1 758 high-quality sequences and 1 365 unigenes including 175 contigs and 1 195 singlets were obtained. These unigenes had 1 283 valid ORFs. COG analysis showed that those proteins coded by EST sequences were divided into 22 classes. Through blast analysis, full length or part cDNA of some genes controlling flower color, development of floral organs, florescence regulation and other breeding value genes were obtained.

本研究以擎天凤梨属品种Ostara为材料,采用SMART技术成功构建了擎天凤梨花器官的质粒型全长cDNA文库,初始文库滴度为3×10^6,插入片段平均长度大于1kb;随机挑取2004个阳性克隆进行5'EST测序,获得高质量序列1758条,经拼接获得1365条单基因簇,其中跨叠群175个和单条序列1195个;经ORF寻找共获得有效ORF1283条;经COG分析EST序列编码的蛋白质被分为22类;经Blast分析后,获得一批花色、花器官发育、花期调控以及其它育种价值基因(包括cDNA全长或片段)。

Secondly, although protein sequence hydropathy prediction result showed ZmDWF1 is an integrated membrane protein, we still successfully expressed the full-length protein fused into the plasmid pET30a , then purified the 70 KD fusion proteins by Ni-NTA agarose affinity chromatography column.

其次,尽管蛋白氨基酸序列的疏水性分析表明,ZmDWF1是一种整合膜蛋白,我们依然将ZmDWF1全长序列克隆到原核表达载体pET30a中,并成功的在大肠杆菌BL21(DE3)中表达了这种预测的膜蛋白全长

Nucleoprotein is also a major immunogenetic antigen for SARS; however, several tests have shown that N protein had the reactivity with other coronavirus positive sera. It is therefore an advantage of S protein as recombinant target antigen for SARS serum diagnosis compared with recombinant N protein. Moreover, S protein has been shown to be the major antigenic determinant that induces neutralizing antibodies and protective immunity to SARS-CoV, so it is also important in designing SARS genetic engineering vaccine.

SARS-CoV S 蛋白由1255 个氨基酸组成,体外表达如此大的蛋白很困难,而且表达蛋白的可溶性不高;其次,实验已经证实,全长S 蛋白含有大量无关的表位,致使表达的全长S蛋白对SARS 阳性血清的检出率低于S 蛋白片段;另外作为疫苗研究,S 蛋白上大量无关表位作为异体蛋白进入体内,会引发机体的过度免疫反应,这也是SARS-CoV 可能的致病机理之一。

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