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201.
Asymmetry in the assembly of the RNAi enzyme complex 总被引:120,自引:0,他引:120
A key step in RNA interference (RNAi) is assembly of the RISC, the protein-siRNA complex that mediates target RNA cleavage. Here, we show that the two strands of an siRNA duplex are not equally eligible for assembly into RISC. Rather, both the absolute and relative stabilities of the base pairs at the 5' ends of the two siRNA strands determine the degree to which each strand participates in the RNAi pathway. siRNA duplexes can be functionally asymmetric, with only one of the two strands able to trigger RNAi. Asymmetry is the hallmark of a related class of small, single-stranded, noncoding RNAs, microRNAs (miRNAs). We suggest that single-stranded miRNAs are initially generated as siRNA-like duplexes whose structures predestine one strand to enter the RISC and the other strand to be destroyed. Thus, the common step of RISC assembly is an unexpected source of asymmetry for both siRNA function and miRNA biogenesis. 相似文献
202.
一株种间融合的三倍体杂种酵母减数分裂产物的遗传学分析 总被引:1,自引:0,他引:1
通过原生质体融合技术将二倍体酿酒酵母(Saccharomycescerevisiaevar.ellopsiodeus)与单倍体糖化酵母(Saccharomycesdiastaticus)构建成遗传上稳定的种间三倍体融合杂种。实验结果表明,这种融合杂种象有性杂种一样能诱导产孢;对其完整和非完整四分子的遗传分析,证明了通过遗传标记互补选择法获得的种间三倍体融合杂种HU-KDF-240在产孢过程中,标记基因发生了分离和交换,出现了亲二型和重组类型;四分子对可溶性淀粉的发酵和不发酵分离比为1∶2,而原养型与营养缺陷型的分离比是1∶1。 相似文献
203.
为了确定从噬菌体抗体文库中筛选出的抗体的属性和方便目的基因的表达及其产物的纯化,对两株具有“1F7”独特型的抗HIV-1gp160抗体基因进行了序列分析并构建了可溶性表达载体.发现3B株含有完整的Fab段,1D株只有重链Fd段.序列测定表明两株克隆的Fd段基因完全相同,其可变区VH属于VHⅠ亚群,而3B株的“轻链”序列与已知的人的κ和λ轻链无同源性.用从另外的Fab抗体文库中筛选出来的3株抗乙肝表面抗原抗体的轻链与3B的重链重组,并选择一个HIV-1gp160特异性较好的重组抗体株,命名为3Bs.构建了1D株与3Bs株的可溶性表达载体,免疫印迹实验证实了具有“1F7”独特型的抗gp160独特型阳性抗体的表达. 相似文献
204.
Chang Liu Jian Tang Yuanmei Ma Xueya Liang Yang Yang Guiqing Peng Qianqian Qi Shibo Jiang Jianrong Li Lanying Du Fang Li 《Journal of virology》2015,89(11):6121-6125
Porcine epidemic diarrhea coronavirus (PEDV) has significantly damaged America''s pork industry. Here we investigate the receptor usage and cell entry of PEDV. PEDV recognizes protein receptor aminopeptidase N from pig and human and sugar coreceptor N-acetylneuraminic acid. Moreover, PEDV infects cells from pig, human, monkey, and bat. These results support the idea of bats as an evolutionary origin for PEDV, implicate PEDV as a potential threat to other species, and suggest antiviral strategies to control its spread. 相似文献
205.
血水草地下部分白屈菜红碱的含量测定 总被引:1,自引:0,他引:1
提出一个单波长扫描测定血水草地下部分白屈菜红碱含量的方法,药材以95%的乙醇提取在320nm处测定,回收率为96.3%(CV=2.39%),结果稳定可靠。 相似文献
206.
247例人乳头瘤病毒的PCR检测结果分析 总被引:1,自引:0,他引:1
247例生殖系分泌物标本用PCR检测人乳头瘤病毒HPV-DNA。结果显示:阳性感染率达87.6%,在与临床疾病密切的亚型中,6、11型感染率(60%),高于16、18型感染率(51.6%),但统计学处理没有显著性差异。女性发病率明显高于男性。易感人群组以20~40岁最高(占感染人数的84.4%),并且有年轻化的趋势(10~20岁年龄组占8.4%)。 相似文献
207.
To reduce the cellobiose inhibition of exoglucanase and endogulcanase and enhance cellulose hydrolysis during simultaneous saccharification and fermentation (SSF), a beta-glucosidase encoding gene named BGL1 was cloned from Saccharomycopsis fibuligera and integrated into the chromosomal rDNA region of the Saccharomyces cerevisiae industrial strain NAN-27 producing NAN-227. Compared with the parental strain, which had no detectable activity, the beta-glucosidase specific activity in NAN-227 was 1.02 IU/mg of protein. When cellobiose was used as the sole carbon source in a shake-flask, NAN-227 consumed 6.2g/L of cellobiose and produced 3.3g/L of ethanol in 48 h. The yield was 0.532 g/g. The parent strain only consumed 1.92 g/L of cellobiose and no ethanol was detected. During the SSF of acid-pretreated corncobs NAN-227 produced 20 g/L of ethanol at 72 h, which was similar to the parent strain when 20IU of beta-glucosidase/g of substrate was added. 相似文献
208.
Lanying Du Xiujuan Zhang Hong Yu Yusen Zhou Shibo Jiang 《Biochemical and biophysical research communications》2010,397(3):580-467
The worldwide outbreak of the swine-origin 2009 H1N1 influenza A virus (IAV) and an increasing number of influenza cases caused by a highly pathogenic avian influenza (HPAI) H5N1 have accelerated the need to develop vaccines and antiviral agents against IAVs. Among various antivirals, neutralizing monoclonal antibodies (mAbs) are considered important passive therapeutics having an immediate effect against viral pathogens. Here we report a pseudovirus neutralization assay for rapid screening of neutralizing mAbs targeting hemagglutinin (HA) of H5N1 and H1N1 IAV. In this study, we generated six pseudoviruses with an HIV-1 backbone, respectively, expressing HA of four clades of H5N1 IAV and the 2009 epidemic H1N1 IAV. The resulting pseudoviruses were able to infect a variety of human and non-human cells, with 293T cells from human kidney as the most susceptible target cells. Using the established pseudovirus neutralization assay, we showed that three of ten selected mAbs specific to HA could potently neutralize infection of a pseudovirus bearing HA from the homologous IAV A/VietNam/1194/2004(H5N1) strain. This was highly consistent with the result of a microneutralization assay testing the same strain of a live IAV. Since the pseudovirus neutralization assay does not involve an infectious virus and can be performed without the requirement of a biosafety-3 laboratory, it may be applied for safe and rapid screening of neutralizing mAbs and antiviral agents targeting HA of IAVs. 相似文献
209.
Ding SJ Wang Y Jacobs JM Qian WJ Yang F Tolmachev AV Du X Wang W Moore RJ Monroe ME Purvine SO Waters K Heibeck TH Adkins JN Camp DG Klemke RL Smith RD 《Journal of proteome research》2008,7(10):4215-4224
Reversible protein phosphorylation is a central cellular regulatory mechanism in modulating protein activity and propagating signals within cellular pathways and networks. Development of more effective methods for the simultaneous identification of phosphorylation sites and quantification of temporal changes in protein phosphorylation could provide important insights into molecular signaling mechanisms in various cellular processes. Here we present an integrated quantitative phosphoproteomics approach and its application for comparative analysis of Cos-7 cells in response to lysophosphatidic acid (LPA) gradient stimulation. The approach combines trypsin-catalyzed (16)O/ (18)O labeling plus (16)O/ (18)O-methanol esterification for quantitation, a macro-immobilized metal-ion affinity chromatography trap for phosphopeptide enrichment, and LC-MS/MS analysis. LC separation and MS/MS are followed by neutral loss-dependent MS/MS/MS for phosphopeptide identification using a linear ion trap (LTQ)-FT mass spectrometer. A variety of phosphorylated proteins were identified and quantified including receptors, kinases, proteins associated with small GTPases, and cytoskeleton proteins. A number of hypothetical proteins were also identified as differentially expressed followed by LPA stimulation, and we have shown evidence of pseudopodia subcellular localization of one of these candidate proteins. These results demonstrate the efficiency of this quantitative phosphoproteomics approach and its application for rapid discovery of phosphorylation events associated with LPA gradient sensing and cell chemotaxis. 相似文献
210.
The relatively slow germination rate of Coniothyrium minitans limits its control efficiency against Sclerotinia sclerotiorum. Pre-germinated conidia of C. minitans enhanced its efficiency significantly: in foliar experiments with oilseed rape, hyphal extension of S. sclerotiorum was inhibited by 68%, while formation of sclerotia was completely inhibited when pre-germinated conidia were applied. 相似文献