Improving siRNA design targeting nucleoprotein gene as antiviral against the Indonesian H5N1 virus     

Risza Hartawan  Dwi Ari Pujianto  Ni Luh Putu Indi Dharmayanti  Amin Soebandrio 《Journal of veterinary science (Suw?n-si, Korea)》2022,23(2)

BackgroundSmall interfering RNA technology has been considered a prospective alternative antiviral treatment using gene silencing against influenza viruses with high mutations rates. On the other hand, there are no reports on its effectiveness against the highly pathogenic avian influenza H5N1 virus isolated from Indonesia.ObjectivesThe main objective of this study was to improve the siRNA design based on the nucleoprotein gene (siRNA-NP) for the Indonesian H5N1 virus.MethodsThe effectiveness of these siRNA-NPs (NP672, NP1433, and NP1469) was analyzed in vitro in Marbin-Darby canine kidney cells.ResultsThe siRNA-NP672 caused the largest decrease in viral production and gene expression at 24, 48, and 72 h post-infection compared to the other siRNA-NPs. Moreover, three serial passages of the H5N1 virus in the presence of siRNA-NP672 did not induce any mutations within the nucleoprotein gene.ConclusionsThese findings suggest that siRNA-NP672 can provide better protection against the Indonesian strain of the H5N1 virus.  相似文献   

新城疫病毒F48E8株cDNA文库的构建   总被引：1，自引：0，他引：1  

陈丽君  吴艳涛 《中国兽医学报》1997,17(2):120-122

以新城疫病毒（ＮＤＶ）中国标准强毒株为材料，分别用基因组ＲＮＡ及ｐｏｌｙ（Ａ）＋ｍＲＮＡ为模板反转录合成ｃＤＮＡ。将ｃＤＮＡ通过同聚物加尾ｐｏｌｙ（ｄＣ）后克隆于末端加有ｐｏｌｙ（ｄＧ）的线性化质粒ｐＧＥＭ－３Ｚｆ（－）上，转化大肠杆菌ＴＧ１，经ＩＰＴＧ和Ｘ－ｇａｌ筛选及电泳检查，其中有５６个克隆含有大小在０．２～２．７ｋｂ范围的外源片段。斑点杂交鉴定有５２个是ＮＤＶｃＤＮＡ克隆，平均长度为１．３１ｋｂ，从而构建了ＮＤＶｃＤＮＡ文库。文库的统计学质量检验表明，拥有５２个克隆的Ｆ４８Ｅ８株ｃＤＮＡ文库可能覆盖ＮＤＶ整个基因组  相似文献   

Effects of intra‐ and interspecific competition and hydropeaking on growth of juvenile Atlantic salmon (Salmo salar)          下载免费PDF全文

Michael Puffer  Ole K. Berg  Ari Huusko  Teppo Vehanen  Sigurd Einum 《Ecology of Freshwater Fish》2017,26(1):99-107

The effects of hydropeaking and intra‐ and interspecific competition on the growth performance (growth in length, mass and lipid content) of juvenile Atlantic salmon Salmo salar and brown trout Salmo trutta were studied in six experimental channels (three experiencing hydropeaking and three controls with a stable discharge of water). Changes in the water‐covered area in the hydropeaking channels were small to avoid fish stranding. Each channel was divided into three similar‐sized sections and stocked with either low or high density of Atlantic salmon, or a mix of Atlantic salmon and brown trout, with the density of the latter equalling the high‐density treatment of Atlantic salmon. A marked effect of competition was visible as salmon in the low‐density treatment were significantly larger (27–33%) and had a higher mass (30–38%) than salmon in both the high‐density salmon treatment and the high‐density salmon and trout treatment. Hydropeaking had only minor and insignificant effects on the growth performance: overall final length, mass and body lipid content in the salmon experiencing hydropeaking differed by ?9%, ?7% and +2% compared with controls. Furthermore, there was no indication that the competitive regime influenced hydropeaking effects. The increase in both intra‐ and interspecific competition among the juvenile salmon had a pronounced and significant effect on growth. Our study adds to the growing evidence that energetic consequences of hydropeaking are likely to be small for Atlantic salmon and that stress and mortality associated with stranding represent the main source of population impact.  相似文献   

Uterine hemodynamics during a short-term progesterone synchronization protocol in ewes     

Beltrame  Renato Travassos  Morais Junior  Nilson Nunes  Careta  Joao Vitor Pagoto  Ribeiro  Ariádna Patricia  Lima  Thales Alves Dutra  da Costa  Ricardo Lopes Dias  Quirino  Celia Raquel 《Tropical animal health and production》2020,52(2):503-509

Tropical Animal Health and Production - Twenty-eight pluriparous and non-lactating Santa Inês sheep were synchronized with vaginal sponge and an intramuscular (IM) injection of...  相似文献   

Effects of earthworms on decomposition and metal availability in contaminated soil: Microcosm studies of populations with different exposure histories     

Tuomas Lukkari  Ari Väisänen 《Soil biology & biochemistry》2006,38(2):359-370

Population-specific differences in the responses of earthworms to simultaneous exposure to Cu and Zn were studied in microcosm experiments. Two populations of Aporrectodea caliginosa tuberculata (Eisen) with different metal exposure histories were chosen for the studies. Microcosms were prepared containing either uncontaminated soil or soils with low or high combined Cu/Zn -concentrations (79/139 or 178/311 mg kg⁻¹ dry mass of soil, respectively). Earthworms from each population were introduced to the microcosm treatments with some microcosms serving as controls without earthworms. One series of microcosms was destructively sampled after 16 weeks incubation in a climate chamber. Survival, growth, reproduction and decomposition by earthworms in each treatment were measured. An additional microcosm series was sampled for soil and earthworm measurements at four weeks intervals to determine temporal changes in the availability of metals in the soils and their accumulation into earthworms. Cu and Zn were sequentially extracted from the soil samples of both microcosm series to estimate mobility and availability of the metals in the soil. Earthworms with long-term exposure history to metal-contaminated soil seemed to tolerate higher soil metal concentrations than earthworms without earlier exposure. Both earthworms and metals affected soil respiration (CO₂ production) and nitrogen mineralization. In addition, earthworms seemed to decrease the mobility and bioavailability of metals in the soil through their burrowing activity.  相似文献   

Nitrogen enrichment causes minimal changes in arbuscular mycorrhizal colonization but shifts community composition—evidence from rDNA data     

Ari Jumpponen  Justin Trowbridge  Keerthi Mandyam  Loretta Johnson 《Biology and Fertility of Soils》2005,41(4):217-224

Intracellular arbuscular mycorrhizal (AM) colonization was compared between nitrogen (NH₄NO₃) fertilized (10 g N m^–2) and nonfertilized tallgrass prairie plots. In the microscopic analyses of host roots, only intracellular coils showed an increasing trend as a result of N fertilization, whereas intracellular colonization by arbuscules, hyphae, or vesicles did not differ between the N treatments. Clone libraries established from pooled PCR products of AM fungi contained exclusively species of Glomus; no other genera were detected indicating that Glomus spp. dominated the host roots. Comparisons between observed and random topologies indicated that cloned sequence placement covaried with N treatment: unique clades within Glomus originated exclusively from N-fertilized or nonfertilized treatments. We conclude that the communities of dominant and most commonly occurring AM fungi changed in response to N amendment, although the root colonization showed minimal or no response.  相似文献   

Local actin polymerization and dynamin recruitment in SV40-induced internalization of caveolae     

Pelkmans L  Püntener D  Helenius A 《Science (New York, N.Y.)》2002,296(5567):535-539

Simian virus 40 (SV40) utilizes endocytosis through caveolae for infectious entry into host cells. We found that after binding to caveolae, virus particles induced transient breakdown of actin stress fibers. Actin was then recruited to virus-loaded caveolae as actin patches that served as sites for actin "tail" formation. Dynamin II was also transiently recruited. These events depended on the presence of cholesterol and on the activation of tyrosine kinases that phosphorylated proteins in caveolae. They were necessary for formation of caveolae-derived endocytic vesicles and for infection of the cell. Thus, caveolar endocytosis is ligand-triggered and involves extensive rearrangement of the actin cytoskeleton.  相似文献   

Protection against potato virus Y (PVY) in the field in potatoes transformed with the PVY P1 Gene     

Tuula Mäki-Valkama  Jari P. T. Valkonen  Ari Lehtinen  Eija Pehu 《American Journal of Potato Research》2001,78(3):209-214

Lines of potato cv. Pito transformed with the P1 gene ofPotato virus Y (PVY^o) in sense or antisense orientation were evaluated for resistance to PVY in the field in 1997 and 1998. The transgenic resistance fully protected the crop from infection with PVY^o transmitted by aphids in both years. These plants were not resistant to the field isolates of the PVY^N strain group, which is in agreement with our greenhouse experiments. Consequently, several transgenic lines produced higher yields than the nontransgenic cv. Pito plants. These results showed that the P1 gene-mediated resistance provides significant benefits under conditions were the incidence of infections and damage by PVY^o are considerable.  相似文献   

Defense-related genes expressed in Norway spruce roots after infection with the root rot pathogen Ceratobasidium bicorne (anamorph: Rhizoctonia sp.)     

Jøhnk N  Hietala AM  Fossdal CG  Collinge DB  Newman MA 《Tree physiology》2005,25(12):1533-1543

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Effect of parent genotype on somatic embryogenesis in Scots pine (Pinus sylvestris)     

Niskanen AM  Lu J  Seitz S  Keinonen K  Von Weissenberg K  Pappinen A 《Tree physiology》2004,24(11):1259-1265

Controlled crosses of seven Scots pine (Pinus sylvestris L.) trees produced 49 families that included both reciprocals and selfings. Embryogenic cultures were initiated from immature megagametophytes and after 6 months in maintenance culture, mature somatic embryos were produced from the surviving 166 lines. The effect of parent genotypes on the cultures was evaluated at initiation of the tissue culture period, after 6 months in maintenance culture and at embryo maturation. The effect of the maternal parent was most pronounced at culture initiation. After 6 months in tissue culture, the maternal effect had decreased and the effects of both parents were significant. By the somatic embryo maturation stage, the maternal effect was still considerable but the paternal effect was no longer detectable. There was little correlation between the ranking of mothers and fathers, indicating that the maternal effect was caused by factors other than the paternal effect. No mother x father interaction was found, indicating that mothers successful at initiation and after 6 months in tissue culture, pollinated by any of the successful fathers, produced somatic lines and mature somatic embryos.  相似文献