Introduction of a point mutation in the KRAS gene of in vitro fertilized porcine zygotes via electroporation of the CRISPR/Cas9 system with single-stranded oligodeoxynucleotides     

Manita Wittayarat  Maki Hirata  Zhao Namula  Yoko Sato  Nhien T. Nguyen  Quynh A. Le  Qingyi Lin  Koki Takebayashi  Fuminori Tanihara  Takeshige Otoi 《Animal Science Journal》2021,92(1):e13534

This study aimed to investigate the efficiency of KRAS gene editing via CRISPR/Cas9 delivery by electroporation and analyzed the effects of the non-homologous end-joining pathway inhibitor Scr7 and single-stranded oligodeoxynucleotide (ssODN) homology arm length on introducing a point mutation in KRAS. Various concentrations (0–2 µM) of Scr7 were evaluated; all concentrations of Scr7 including 0 µM resulted in the generation of blastocysts with a point mutation and the wild-type sequence or indels. No significant differences in the blastocyst formation rates of electroporated zygotes were observed among ssODN homology arm lengths, irrespective of the gRNA (gRNA1 and gRNA2). The proportion of blastocysts carrying a point mutation with or without the wild-type sequence and indels was significantly higher in the ssODN20 group (i.e., the group with a ssODN homology arm of 20 bp) than in the ssODN60 group (gRNA1: 25.7% vs. 5.4% and gRNA2: 45.5% vs. 5.9%, p < .05). In conclusion, the CRISPR/Cas9 delivery with ssODN via electroporation is feasible for the generation of point mutations in porcine embryos. Further studies are required to improve the efficiency and accuracy of the homology-directed repair.  相似文献   

外源肌苷酸对种鸡产蛋性能、蛋品质及种蛋孵化性能的影响     

牛自康  吴乐  张鑫蕊  叶成智  宦海琳  张峰  魏明星  闫俊书 《中国畜牧兽医》2021,48(1):172-177

本试验通过在种鸡的基础饲粮中加入肌苷酸（IMP）来研究其对种鸡产蛋性能、蛋品质和种蛋孵化性能的影响。选择遗传背景相同、体重相近,产蛋率达到5%的20周龄健康AA种鸡864羽,随机分为2组（对照组饲喂基础饲粮;试验组饲喂基础饲粮＋0.5%肌苷酸）,每组6个重复,每个重复72羽,试验期30 d。测定试验期间所有种蛋的平均蛋重、产蛋率和合格蛋率;每组随机选取部分种蛋进行蛋品质测定;于孵化开始第17天,通过照蛋处理统计种蛋受精率,第21天出雏时,计算受精蛋孵化率、入孵蛋孵化率和健雏率。试验结果表明：与对照组相比,试验组种鸡的平均蛋重、产蛋率和合格蛋率均有升高的趋势,但差异不显著（P>0.05）;蛋形指数显著升高（P<0.05）,蛋壳强度、蛋壳厚度和蛋黄颜色显著降低（P<0.05）;受精率、受精蛋孵化率和入孵蛋孵化率显著升高（P<0.05）,健雏率和雏鸡体重有升高的趋势,但差异不显著（P>0.05）。综上,0.5%外源肌苷酸对种鸡的产蛋性能和种蛋孵化性能均有促进作用。  相似文献   

实例获取网络在车辆诊断专家系统中的应用     

张代胜  王悦  陈朝阳  张乐  徐进 《农业机械学报》2002,33(6):29-33

针对基于规则推理和模型推理专家系统的局限性，提出将实例获取网络应用于基于实例推理的车辆诊断专家系统中，介绍了实例获取网络在车辆诊断专家系统中的设计方法、工作过程及其算法的改进方案，探讨了实例获取网络、回收机制及相关策略的实现方法，对基于实例推理专家系统设计策略、实施步骤和组织结构上的一些共性问题做了详述。  相似文献   

基于废弃物的潞安煤矿废弃地改良土壤基质配比研究   总被引：2，自引：0，他引：2  

王乐  郭小平  韩祖光  邓川  王川  曾旸  李峰  程冀南 《土壤》2020,52(1):145-152

为解决潞安矿区煤矸石山、塌陷地生态修复缺土少肥问题,本研究将粉煤灰、污泥与垃圾堆肥以5%、10%、20%体积比例正交混合配制改良土壤基质进行盆栽试验,观测不同配比土壤的理化性质、养分及重金属含量、高羊茅与紫叶小檗生长状况,并用主成分–聚类分析法筛选最优配比。结果表明:添加垃圾堆肥可以提高土壤有效养分与有机质含量,对土壤理化性质改良有明显效果;添加污泥仅提升土壤有效磷含量;添加粉煤灰在降低土壤容重、增大总孔隙度与非毛管孔隙度上效果明显,但对土壤p H、阳离子交换量(CEC)与碱解氮的改良具有显著负效应。各废弃物改良基质的碱解氮、有效磷、速效钾、有机质等含量均较高,土壤重金属含量也处在安全范围,而土壤容重、非毛管孔隙、pH、电导率(EC)与CEC等指标性质较优的处理组为粉煤灰∶污泥∶垃圾堆肥∶土=5%∶20%∶20%∶55%、10%∶10%∶20%∶60%、20%∶5%∶20%∶55%3个处理。经过综合筛选,本研究基质最优混合配比为粉煤灰∶污泥∶垃圾堆肥∶土=5%∶20%∶20%∶55%,可作为当地矿区废弃地生态修复客土材料推荐方案。  相似文献   

考虑收缩的爆炒热/质传递过程模拟与验证     

谢乐  邓力  李静鹏  曾雪峰  闫勇  石宇  苏婕妤 《农业工程学报》2020,36(18):251-262

为研究爆炒中食品多孔介质热/质传递机制及烹饪成熟和品质变化规律，考虑收缩-水分损失关系，基于多孔介质理论，结合傅里叶定律、牛顿冷却定律和达西定律，构建了爆炒中有蒸发、考虑收缩的食品含湿非饱和多孔介质热/质传递数学模型，开展了爆炒数值模拟。考虑收缩后模拟值与实测温度历史（Least Summation of the squared Temperature Difference for overall target，LSTD=4.40 ℃）、平均含水率（LSTD=1.42%）和体积收缩率（LSTD=1.05%）吻合更为良好。模拟分析了爆炒热/质传递对颗粒表面蒸发、收缩、内部压力、水分和温度分布的影响机理及火候控制手段的作用，结果表明：爆炒强对流传热使颗粒蒸发剧烈造成水分损失；收缩主要由水分损失引起，可以增大传热效率并影响颗粒内部压力变化，由成熟值理论研判，蒸发收缩对烹饪成熟起到促进作用，并有利于提高烹饪品质，是爆炒技术优势的一部分；火候控制手段通过改变颗粒特征尺寸、能量传递速率和流体-颗粒的换热时间/接触面积对颗粒成熟时间和含水率等烹饪品质产生极显著（P<0.01）影响。  相似文献   

不同种植年限土壤微量元素对山银花品质的影响   总被引：1，自引：0，他引：1       下载免费PDF全文

颜秋晓  林昌虎  高安勤  张珍明  乐乐  何腾兵 《水土保持通报》2015,35(3):299-304

[目的]探索不同种植年限土壤微量元素对山银花品质的影响,筛选山银花高产优质的最佳种植年限范围。[方法]采用野外调查和室内分析结合的方法,研究贵州省绥阳县不同种植年限山银花土壤微量元素对山银花品质的影响。[结果]不同种植年限下土壤pH值为4.67~6.01,土壤微量元素平均含量为:Cu 17.17~37.50 mg/kg,Mo 1.52~2.24 mg/kg,Mn 444.6~544.26 mg/kg,Zn 102.06~114.39mg/kg;山银花花蕾微量元素平均含量为:Cu 6.91~14.52 mg/kg,Mo 0.16~0.24 mg/kg,Mn 30.06~79.36mg/kg,Zn 12.73~23.54mg/kg;不同种植年限山银花绿原酸含量为2.988%~7.840%。[结论]土壤中Zn,Cu含量随种植年限增加而递增,Mo,Mn含量总体呈下降趋势。11~15a的山银花绿原酸含量最高,植物生长状况良好,花蕾微量元素含量总体随年限增加而增加。微量元素Cu,Zn能促进山银花的生长和绿原酸的合成,Mn元素与山银花的茎粗和叶片生长有关。  相似文献   

Aggregate stability and microbial community dynamics under drying-wetting cycles in a silt loam soil     

Diego Cosentino  Claire Chenu  Yves Le Bissonnais 《Soil biology & biochemistry》2006,38(8):2053-2062

Aggregate stability often exhibits a large inter-annual and seasonal variability which occurs regardless of residue treatments and is often larger than the differences between soils or cropping systems. Variations in soil moisture and seasonal stimulation of microbial activity are frequently cited as the major causes. The goal of this paper was to evaluate the effects of drying-rewetting cycles on aggregate stability and on its main microbially mediated agents from a mechanistic point of view. The 3-5 mm aggregates of a silty soil were incubated at 20 °C for 63 days with the following treatments and their combinations: (i) with or without straw input and (ii) with or without exposure to four dry-wet cycles. Microbial activity was followed by measuring the soil respiration. We estimated the microbial agents of aggregate stability measuring hot-water extractable carbohydrate-C, microbial biomass carbon and ergosterol content. We measured the water drop penetration time to estimate the hydrophobicity and aggregate stability according to Le Bissonnais [1996. Aggregate stability and assessment of soil crustability and erodibility: I. Theory and methodology. European Journal of Soil Science 47, 425-437] to distinguish three breakdown mechanisms: slaking, mechanical breakdown and microcracking. The addition of straw stimulated microbial activity and increased the resistance to the three tests of aggregate stability, enhancing the internal cohesion and hydrophobicity of aggregates. All the estimated microbial agents of aggregate stability responded positively to the addition of organic matter and were highly correlated with aggregate stability. Fungal biomass correlated better with aggregate stability than total microbial biomass did, showing the prominent role of fungi by its triple contribution: physical entanglement, production of extracellular polysaccharides and of hydrophobic substances. Dry-wet cycles had less impact on aggregate stability than the addition of straw, but their effects were more pronounced when microbial activity was stimulated demonstrating a positive interaction.  相似文献   

A new approach to examine the relationships between sensory and gas chromatography-olfactometry data using Generalized Procrustes analysis applied to six French Chardonnay wines     

Le Fur Y  Mercurio V  Moio L  Blanquet J  Meunier JM 《Journal of agricultural and food chemistry》2003,51(2):443-452

  相似文献   

Synthesis of haptens and conjugates for ELISAs of phytoestrogens. Development of the immunological tests     

Bennetau-Pelissero C  Le Houérou C  Lamothe V  Le Menn F  Babin P  Bennetau B 《Journal of agricultural and food chemistry》2000,48(2):305-311

Seven carboxylic acid haptens of isoflavonoids were synthesized, with the spacer arm on the oxygen atom at the C7 position for one series, with formononetin, daidzein, equol, biochanin A, and genistein, and at the C8 position for a second series, with only formononetin and daidzein. The different haptens were coupled to bovine serum albumin (BSA) and to swine thyroglobulin (Thyr). Polyclonal antibodies were generated against the BSA conjugates. Enzyme-linked immunosorbent assays (ELISAs) were developed based on competition between free phytoestrogens and the Thyr-hapten conjugates for specific antibodies. IC(50) values of the standard curves ranged between 0.8 and 20 ng/mL that is, 0.3 and 9.2 pmol/well. The antibodies obtained should be useful for assays in vegetable matter as well as in biological fluids after a separation step. These ELISAs should be valuable also in the food industry to control phytoestrogen concentrations prior to and after processing.  相似文献   

Bootstrap model selection in generalized linear models     

Wei Pan  Chap T. Le 《Journal of Agricultural, Biological & Environmental Statistics》2001,6(1):49-61

Model selection is a central component of data analysis. Though there are a variety of methods for likelihood-based estimation methods, there are relatively few for non-likelihood-based generalized linear models (GLMs), such as in the quasi-likelihood and generalized estimating equation (GEE) approaches. In this paper, we develop basic and bias-corrected bootstrap approaches to estimate the predictive mean squared error (PMSE) of a model and use the PMSE for model selection. Simulation studies show that the bias-corrected bootstrap estimate works well when quasi-likelihood or GEE is used to fit either overdispersed or correlated response GLMs. For correlated response data, when the marginal distribution assumption is (almost) correct, Akaike’s information criterion (AIC) and Bayesian information criterion (BIC) calculated under the working independence model also perform well. For illustration, the methods are applied to data sets from evolutionary biology and teratology.  相似文献