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blast分析
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  blast analysis
trichocarpa (AF057708) determined by blast analysis in the GenBank.
      
A BLAST analysis revealed an open reading frame (ORF) that appears to encode for the Tetrahymena DNA-[adenine] methyltransferase ((MTase), based on the presence of motifs characteristic of the enzymes in prokaryotes.
      
In silico analysis indicated that the coding region contains a single intron and translates into an 893 amino acid protein, with BLAST analysis identifying five conserved nitrate reductase domains within the protein.
      
The bacterial strains were identified as Bacillus species by PCR amplification of the 16S rRNA gene, BLAST analysis, and comparison with sequences in the GenBank nucleotide database.
      
The BLAST analysis demonstrated that the cloned sequence is 100% homologous with sequences reported overseas.
      
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RNAs of the barks and latexes from different rubber trees(Hevea brasiliensis)cultivated in the experimental fields of Chinese Academy of Tropical Agricultural Sciences were isolated for mRNA differential display analysis.Screened by4OligodT 12 (C,A,G)GC and ten26-oligonucleotide arbi trary primers,differential cDNA fragments-Hb 1(725bp),Hb 2,Hb 3and Hb 4,only appeared in the barks and latexes of healthy trees of three clones of Hevea brasiliensis(RRIM600,RY13-28,RY1-20).Hb 1was isolated and purified.It...

RNAs of the barks and latexes from different rubber trees(Hevea brasiliensis)cultivated in the experimental fields of Chinese Academy of Tropical Agricultural Sciences were isolated for mRNA differential display analysis.Screened by4OligodT 12 (C,A,G)GC and ten26-oligonucleotide arbi trary primers,differential cDNA fragments-Hb 1(725bp),Hb 2,Hb 3and Hb 4,only appeared in the barks and latexes of healthy trees of three clones of Hevea brasiliensis(RRIM600,RY13-28,RY1-20).Hb 1was isolated and purified.It was confirmed by RNA dot hybridization that Hb 1was ex pressed in latexes of rubber trees,but its expression decreased significantly in latexes of TPD(tap ping panel dryness)trees,which showed that Hb 1was correlated with TPD.Hb 1was cloned and se quenced.After searched in Genbank,no similar sequences were reported.This cDNA may provide basis for obtaining integral genes associated with TPD and studying the molecular mechanism underlying TPD.

从中国热带农业科学院橡胶树早期预测试验地采集不同品系的橡胶树胶乳和树皮组织,分别提取纯化mRNA合成cDNA第一链。通过4个简并锚定引物和10个随机引物进行PCR扩增,经6%变性聚丙烯酰胺凝胶电泳后发现了至少4个差异DNA片段只在健康树中表达而在死皮树中没有出现。这4个差异DNA片段分别命名为Hb1(725bp),Hb2,Hb3,Hb4。对Hb1进行了分离、纯化,经Northern杂交证实,Hb1在健康树胶乳中表达活跃,而在死皮树胶乳中表达受到强烈抑制,表明Hb1是与死皮病相关的cDNA片段。进一步将Hb1片段进行回收和克隆,并进行DNA序列分析。序列在GenBank中进行BLAST分析未发现相关同源片段。死皮病相关cDNA片段的获得,将为分离全长死皮病相关基因,搞清该基因表达调控的机理提供条件,进而为从分子水平上阐明死皮病的致病机理打下基础。

Using a universal primer of family Potyviridae, potato virus A (PVA) was identified from potatoes in Hangzhou city. One thousand six hundred and eighty seven nucleotide sequence of 3' terminal genome was determined, including NIb, CP and 3' UTR. DAG motif, which was closely related to the aphid transmission, was identified at the position 215 217aa in polyprotein. Comparison to other 19 isolates of PVA suggested that Hangzhou isolate was most similar to Alc isolate from the Netherlands, and their homology...

Using a universal primer of family Potyviridae, potato virus A (PVA) was identified from potatoes in Hangzhou city. One thousand six hundred and eighty seven nucleotide sequence of 3' terminal genome was determined, including NIb, CP and 3' UTR. DAG motif, which was closely related to the aphid transmission, was identified at the position 215 217aa in polyprotein. Comparison to other 19 isolates of PVA suggested that Hangzhou isolate was most similar to Alc isolate from the Netherlands, and their homology of amino acid sequence of CP was 99.6%. Phylogenetic tree analysis confirmed that some significant groups of PVA isolates existed.

采用马铃薯Y病毒属通用引物Sprimer扩增了引起浙江杭州郊区发生的马铃薯病毒基因组 3’ 末端 16 87核苷酸序列 ,它包括NIb ,CP和 3’ UTR ,3’ 末端含Poly(A)尾 ,蚜传基序DAG位于多聚蛋白第 2 15~ 2 17氨基酸。BLAST分析表明该病毒为马铃薯A病毒 (PVA) ,它与已报道的 19个PVA分离物CP氨基酸进行比较 ,PVA浙江分离物与荷兰Alc分离物同源性最高 ,为 99.6 %。系统进化树分析表明 ,PVA分离物间存在一些明显的进化群体。

A cosmid library of Xenorhabdus nematophilus BP, the symbiotic bacteri a of entomopathogenic nematode (EN) Steinernema carpocapsae, was constructed and two clones of the resulting library, cos76 and cos83 were found possess str ong and moderate oral toxicity against neonate of cotton ball worm Helicoverpa arm igera, respectively. To identify the toxin genes presented in these two clones , seven pairs of primers were designed based on the published sequences and used for PCR amplification with both...

A cosmid library of Xenorhabdus nematophilus BP, the symbiotic bacteri a of entomopathogenic nematode (EN) Steinernema carpocapsae, was constructed and two clones of the resulting library, cos76 and cos83 were found possess str ong and moderate oral toxicity against neonate of cotton ball worm Helicoverpa arm igera, respectively. To identify the toxin genes presented in these two clones , seven pairs of primers were designed based on the published sequences and used for PCR amplification with both clones. All seven pairs of primers yielded prod ucts from cos83 while only five did from cos76. All PCR products were sequenced. The results showed that each sequence of the five PCR products obtained from c os76 was identical with that from cos83, indicating that the toxin genes in cos 76 were part of those in cos83. The BLAST results showed that the amino acid seq uences deduced from seven PCR products from cos83 had mean homology of 95% and 65% with those from insecticidal toxin genes of X.nematophilus PMF1296 and P.luminescens W14, respectively. The results indicate that toxin produced by cos83 is a member of the oral toxin family of EN symbiotic bacteria but it diff er substantially from other members of the family therefore worthy of further i nvestigation.

构建了昆虫病原线虫共生菌XenorhabdusnematophilusBP品系的粘粒文库并用生物测定的方法从中筛选到 2个对棉铃虫初孵幼虫有口服抑杀作用的克隆cos83和cos76。为初步确定这两个克隆的毒素基因与已报道基因的差异程度 ,根据已发表的共生菌毒素基因序列资料设计了 7对引物对这两个克隆进行PCR扩增并对扩增产物进行测序和分析。从毒力较强的cos83中 ,7对引物均扩增出与目标片段大小一致的产物 ,而从cos76中只有 5对扩增到目标片段。测序结果显示 ,cos76的 5个PCR扩增产物与cos83的对应扩增产物DAN序列同源性为 10 0 %。以cos83的 7个PCR扩增产物所编码氨基酸序列进行BLAST分析 ,它们与X .nematophilusPMF12 96和PhotorhabdusluminencencW14毒素相应区间的平均同源性分别为 94%和 5 8% ,说明cos83毒素是共生菌口服毒素家族的一员但与同类的其它毒素有一定的差异 ,值得对其杀虫谱 ,作用机理等进行进一步的研究。

 
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