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After thecomplete genome extraction of the strain was performed, the genomic DNA was partiallydigested by restriction enzyme Sau3AⅠ, the DNA fragments from 1 to 5Kb was clonedinto prokaryote expression vector pET-28a-c, and transformed host bacteria. The resultsshowed that we succeeded in constructing the gene expression library of haemophilusparasuis serovar 5, which is fundamental for the study of advanced gene screening. Inaddition, primer design was performed based on haemophilus influenzae in this study. In addition, PCR was performed by using genomic DNA of haemophilus parasuisserovar 5 as the template. The results demonstrated that we obtained two neo-gene:23SrRNA gene(conserved gene belonging to the large-subunit of ribosome) and adenylatecyclase gene(encodes adenylate cyclase and participates in converting adenyl nucleosidetriphosphate to cyclic adenosine3",5"-monophosphate). Furthermore, the phylogeneticanalyses between the species was performed, and neighbor-joining tree was constructedbased on comparison of 23S rRNA gene sequences, so it was illuminated betweenHaemophilus parasuis and other species in molecular evolution relationship.

选择我国流行优势菌株副猪嗜血杆菌血清5型地方株为研究对象,提取细菌基因组DNA,用限制性内切酶Sau3AⅠ对基因组DNA进行部分酶切,回收大小为1~5Kb的DNA片段,将其连接入原核表达载体pET-28a-c,最后转化宿主菌,结果成功地构建了基因表达文库,为后续的基因筛选工作奠定基础;另外,本研究选择嗜血杆菌属的流感嗜血杆菌为参考对象进行引物的设计,以副猪嗜血杆菌血清5型地方菌株的基因组DNA为模板,进行PCR扩增反应,结果表明成功地获得两个新基因:23S rRNA基因(存在于核糖体大亚基中的保守性基因)和腺苷酸环化酶基因(负责将腺嘌呤核苷三磷酸转变为环腺苷酸),并进一步做了不同物种之间的分子系统发育分析,构建了基于23S rRNA基因的邻接法系统发育树,阐明了副猪嗜血杆菌与其它菌种的分子进化关系。

We summarized the progress in research on avirulence genes of the rice blast fungus, including the importance of the research, avirulence genes cloned and their interaction characteristics with rice resistance genes.

本文从研究稻瘟菌无毒基因的意义、已鉴定和克隆的稻瘟菌无毒基因、稻瘟菌无毒基因与其抗病基因的互作特点等几个方面,对稻瘟菌无毒基因研究进展作了简要评述。

The total RNA was abstracted from the larvae of Boophilus microplus, and a 1982bp Bm86 gene was amplified by RT-PCR. The target gene was subcloned into T vector. The sequencing showed that the nucleotide sequence of the cloned Bm86 gene shared 97.4% homology with the data published in and this fragment contained the complete open reading frame of Bm86 gene.

为了克隆微小牛蜱Bm86 基因及构建该基因的表达载体,以微小牛蜱饥饿幼蜱的破解物提取的总RNA为模板,参照已发表的微小牛蜱Bm86基因的核苷酸序列,设计了1对引物,采用RT-PCR技术获得微小牛蜱Bm86基因;将Bm86基因克隆入载体,并进行序列分析,结果证明,克隆的Bm86基因序列与GenBank上登录的Bm86基因序列的同源性达97.4%,而且该序列包含完整的开放阅读框。

ErmE derived from Saccharopolyspora erythraea, which encodes a ribosome dimethyltransferase, was expressed in S. thermotolerans, to increase the substance resistance during the biotransformation of tylosin to AIV.

研究表明acyB2基因应该是碳霉素生物合成基因簇中的一个全局性正调控因子,它不仅负责对acyB1基因的正调控作用,而且还至少负责对acyA基因和红霉糖糖基转移酶基因的正调控。

Based on these observations, we predict that the transmission of GA-signaling genes occurs in a sporophytic manner, since the protein products and/or mRNA transcripts of these genes may be introduced into pollen-carrying mutant alleles, whereas GA synthesis genes are transmitted in a gametophytic manner, since these genes are preferentially expressed after meiosis.

基于这些观察,我们预测该遗传算法,基因信号传输时,在孢子的方式,因为蛋白质的产品和/或基因这些基因的转录可以引入花粉携带突变基因,而合成的基因遗传传递以配子体的态度,因为这些基因是优先减数分裂后表示。

Drug sensitive test and three-dimensional test220 strains of Pa were isolated from hospitalized patients between 2003 and 2007. K-B method was used to tested the susceptibility of 10 different antibiotics. IRPa was screened by testing the minimal inhibitory concentration of imipemem by using agar diluiion method.The susceptibility of these IRPa to the antibiotics was analysised. Three-dimensional test was used to identify the different kinds of beta lactamases from 220 strains of Pa.2.Carbarpenems hydrolytic enzyme genes and oprD2 gene were detectedamong the selected IRPa strains, PCR method was performed to detect carbapenemase genes which included GES、KPC、SPM、VIM、IMP、GIM gene and the oprD2 gene;Multiplex PCR were used to detect OXA genes and plasmid-mediated AmpC beta lactamase genes; The expression of the chromosomal AmpC beta lactamases and oprD2 genes in IRPa strains were analyzed by Real-time PCR.3.Identification and characterization of integronsIntegrase gene was detected by PCR, and the classification of integrons was performed by using restriction fragment length polymorphism.PCR was performed to detect the qacE△1-sull gene,and the gene cassetes which are located at variable region of integrons in the strains were detected to be positive.

方法1、药敏实验和三维实验收集2003~2007年临床分离的220株Pa,对这些菌株采用K-B法测定10种临床常用抗生素的药敏情况,同时采用琼脂稀释法检测亚胺培南的最低抑菌浓度(Minimal inhibitory concentration,MIC),筛选出对亚胺培南耐药的铜绿假单胞菌,并分析其对其它抗生素的药物敏感率;采用三维实验的方法分析220株Pa产β内酰胺酶的类型。2、碳青霉烯类水解酶和oprD2蛋白的检测针对鉴定的IRPa菌株,采用普通PCR方法检测具有碳青霉烯水解作用的β内酰胺酶耐药基因(GES、KPC、SPM、VIM、IMP、GIM基因)和oprD2基因,采用多重PCR的方法检测OXA型基因和质粒携带的AmpC酶基因,用荧光定量RT-PCR方法检测oprD2蛋白基因表达情况;同时对产AmpC酶的Pa(25株,含IMP耐药和敏感株)用RT-PCR方法检测AmpC酶基因的表达量情况。3。

Plant traits were categorized to three classes: the first class traits were dominated by major gene with little genetic effect of polygene, including days from sowing to flowering, days from sowing to boll opening, lint percent, days from squaring to flowering, days from flowering to boll opening and plant bolls. The hereditability of major gene and polygene were about 36. 85%~66. 79% and 2. 83%~12. 64%, respectively.

通过对短季棉早熟及其相关性状的主-多基因综合分析发现:主基因普遍存在,各时期至少有一对主基因起主要作用,同时受多基因的修饰,在短季棉发育的不同时期,其主基因的对数、作用效果、方向以及主、多基因的遗传率均不同。

Was simultaneously injected into cisterna magna at the second injection blood in the presence of subarachnoid blood. At 2 to 4 days, GFP was expressed in leptomeninges over the brain stem, cortex and cerebral arteries, smooth muscle cells of small vessels were occasionlly transduced. GFP was expressed in adventitia of spastic basilar artery on days 2 (day 5 after first injection blood) after injection 〓, but was undetectable by days 4, transgene was not expressed in medial or intimal layers. The prensence of subarachnoid blood can not prevent access of adenovirus to vessels and transgene expression.

注入腺病毒载体后2天(初次注血第5天)行荧光显微镜、免疫组织化学和RT-PCR检测,结果在颅内大血管如基底动脉的外膜可见外源基因的表达,而外源基因不能转移至血管中膜和内膜,4天时(初次注血第7天)基底动脉的外膜中外源基因表达消失;注入腺病毒载体后2~4天,外源基因可以有效转移至颅底软脑膜细胞,小血管外膜和平滑肌层也可见外源基因的表达,表明蛛网膜下腔内的血凝块不能阻止腺病毒载体介导外源基因转移至颅内血管。

Majority of acute leukemias in infant, either acute lymphoblastic leukemia or acute myeloblastic leukemia, posses a chromosomal translocation affecting the 11q23 chromosome region which specifically inoles the mixed-lineage leukemia gene.1-3 Most pediatric leukemias with MLL rearrangement clearly hae a remarkably short latency.1,4 MLL gene rearrangement is also associated with secondary leukemias of patients preiously treated with the topoisomerase II inhibitors.4 The latency of these secondary leukemias is similarly ery short.4 Of note, the concordance rate of leukemia with MLL rearrangement in infant monozygotic twins approximates to 100%,1,4 and identical breakpoint in the MLL gene was shared in these pairs of identical twin infants with concordant ALL.1,4 Moreoer, the unique and clonotypic MLL fusion gene was detectable in neonatal blood spots for Guthrie cards from non-twined indiiduals who subsequently deeloped ALL.1,4 These obserations indicate not only that MLL fusion is generated in utero but also that MLL fusion proteins could be capable of inducing leukemic transformation with few, if any, secondary mutations.2,3,4 Greaes et al speculate that an MLL fusion protein somehow promotes rapid transition to full-blown disease in patients ia ery rapid clonal expansion, genetic instability, or inhibition of DNA damage repair.4 In general, for clonal expansion of malignancies, tumor cells often hae acquired strategies that escape immune sureillance of the hosts.5,6 Immune escape mechanisms also contribute to the failure of graft-ersus-leukemia effect after allogeneic hematopoietic stem cell transplantation.7 Therefore, leukemia cells could acquire some immune escape mechanisms during leukemogenesis.

绪论 绝大多数的婴儿白血病,不管是急性淋巴性白血病或是急性骨髓性白血病,在染色体11q23部位有染色体易位的情况;这个部位的染色体易位牵连了混合谱系白血病基因。大多数具有MLL基因重排的儿童白血病潜伏期明显短很多。MLL基因重排也和经拓扑异构酶II抑制剂治疗后的继发性白血病有关。这些继发性白血病的潜伏期类似地都非常的短。很重要的是,单卵双胞胎婴儿同时患有或同时免于MLL基因重排阳性的白血病的一致性接近100%;并且同样患有ALL的同卵双胞胎的MLL基因的断裂点是一致的。而且,这种独特的克隆特异性的MLL融合基因能够从那些得ALL的非双生个体出生时的血斑标本中检测到。这些发现表明MLL融合基因产生在胎儿还在子宫的是后,而且MLL融合蛋白能过和其他的基因突变一起诱导白血病的产生。Greaes 等推测MLL融合蛋白在某种情况下同过快速克隆增殖,遗传的不稳定性或是DNA损伤修复的抑制促使疾病迅速地全面爆发。恶性肿瘤细胞的克隆增殖通常已经获得了逃避机体免疫监视的能力。免疫逃避机制也归因于异体外周血干细胞移植后移植物抗白血病作用的失效。所以,白血病细胞在白血病的产生过程中可能获得了某些免疫逃脱机制。

The plasmid pDM803, carrying the bar gene, conferring resistance to the herbicide, and the screenable uidA gene, was transferred into wheat by particle bombardment.

在此基础上,将含有bar基因和GUS报告基因的质粒pDM803用基因枪法转化小麦,获得可育的转基因小麦植株,从而建立了适于中国春性小麦基因型的基因枪转化体系。

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