伪狂犬病病毒在NF-κB家族p65基因敲除细胞系的复制规律研究     

常雯茹  段利芳  杨乐  马英先  王棋  翟云云  周鲁豫  张爽  明胜利  杨国宇  王江  褚贝贝 《中国畜牧兽医》2021,48(1):83-92

试验旨在研究伪狂犬病病毒（PRV）在NF-κB家族p65基因敲除细胞系中的复制规律。利用慢病毒介导的CRISPR/Cas9基因定点修饰技术构建猪肺泡巨噬细胞（3D4/21）p65基因稳定敲除细胞系。通过构建p65-sgRNA重组质粒,转染至HEK293T/17细胞,收取慢病毒,感染3D4/21细胞后利用嘌呤霉素筛选获得多克隆细胞系,T7核酸酶检测敲除效率,再通过有限稀释法获得3D4/21-p65^-/-的稳定细胞系。CCK-8试剂盒检测3D4/21细胞中敲除p65基因后对细胞增殖的影响;流式细胞术检测PRV-GFP感染3D4/21及3D4/21-p65^-/-细胞后病毒增殖的差异;实时定量PCR检测PRV感染3D4/21及3D4/21-p65^-/-细胞后PRV gB、TK基因mRNA表达水平及PRV感染细胞诱导的IL-1β和IL-6基因mRNA水平表达的变化;Western blotting检测PRV-QXX感染3D4/21及3D4/21-p65^-/-细胞后PRV gB、gE蛋白的表达;滴度测定检测PRV-QXX感染3D4/21及3D4/21-p65^-/-细胞后子代病毒滴度。结果表明,sgRNA2和sgRNA3的基因编辑效率较高,对其进行克隆化培养进而获得敲除p65基因的稳定表达细胞系;CCK-8试剂盒检测细胞活力表明,p65基因敲除对细胞活力无影响;流式细胞仪检测表明,同一时间点PRV-GFP在3D4/21-p65^-/-中的增殖显著高于对照细胞;实时荧光定量PCR表明在3D4/21细胞中敲除p65基因促进了PRV gB、TK基因的mRNA表达水平,而抑制了IL-1β、IL-6基因的mRNA表达;Western blotting结果表明,在3D4/21细胞中敲除p65基因促进了PRV gB、gE蛋白的表达;滴度测定结果表明,同一时间点PRV-QXX在3D4/21-p65^-/-细胞中子代病毒的复制显著高于对照细胞。以上结果均表明,p65基因敲除可促进PRV在3D4/21细胞中复制。  相似文献   

Effects of a Dual‐Frequency Frequency‐Sweeping Ultrasound Treatment on the Properties and Structure of the Zein Protein     

Xiaofeng Ren  Xun Wei  Haile Ma  Huiji Zhou  Jingjing Guo  Shuyun Mao  An Hu 《Cereal Chemistry》2015,92(2):193-197

We investigated the effects of a dual‐frequency frequency‐sweeping ultrasound (DFFSU) treatment on the functional properties and structure of zein. The solubility of ultrasound‐treated zein proteins increased slightly but significantly as the treatment time increased. The results showed that the DFFSU treatment had an obvious influence on the mean particle size and the size distribution. A significant (P < 0.05) increase in the size of the particles with respect to time was observed after a sonication time of more than 20 min in zein solutions. Differential scanning calorimetry results showed that sonication alters the thermal behavior of zein. Circular dichroism spectra showed a small increase in the percentage of ordered structure elements within the protein molecule. After 60 min of ultrasonication, the percentage of α‐helix structures increased by 0.9%, whereas the percentage of β‐sheets and β‐turns decreased by 0.5%. Microstructural analyses by scanning electron microscopy showed that several microholes appeared in the zein following ultrasonic pretreatment. Under the conditions investigated in this study, DFFSU treatment was found to affect the studied functional properties of the zein protein. This technology could be used to obtain improved functional properties in some protein samples.  相似文献   

干旱胁迫对3种冷季型草坪草光谱反射率及生理特征的影响     

赵志军  单贵莲  段新慧  姜华  任健  陈功  初晓辉 《草原与草坪》2016,(6):23-29

为探讨高光谱分析技术在草坪草水分监测上的应用,以草地早熟禾、高羊茅、多年生黑麦草为试验材料,测定了干旱胁迫下3种冷季型草坪草生理指标及光谱反射率。结果表明:随着干旱胁迫程度的加深,供试草坪草叶片相对含水量和叶绿素含量显著下降(P0.05),丙二醛含量、脯氨酸含量及敏感波段的(550和760 nm)的光谱反射率显著上升(P0.05),3种草坪草抗旱性能的强弱顺序为:高羊茅多年生黑麦草草地早熟禾。3种草坪草敏感波段光谱反射率(R_(550 nm),R_(760 nm))及高光谱参数(Rg,R0,GNDVI)与生理指标间呈显著或极显著相关,以R_(550 nm)与生理指标间的相关系数绝对值最大。同时R_(550 nm)构建了供试草坪草生理指标的估测模型,经检验,模型估测值与实测值相关性达显著或极显著水平。  相似文献   

热水处理对鲜切石榴籽粒贮藏品质、抗氧化能力及微生物变化的影响     

王琼  初丽君  王敏  寇莉萍 《北方园艺》2016,(1):106-110

以石榴为试材,采用热处理法,研究了不同处理条件对保鲜期间鲜切石榴籽粒贮藏品质、抗氧化能力及微生物变化的影响。结果表明:鲜切石榴籽粒经过40℃10min、45℃6min、50℃4min的热处理,其中50℃4min的热处理能够提高石榴籽粒可溶性固形物含量,减轻石榴籽粒贮藏期间的质量损失,提高石榴籽粒的抗氧化活性,抑制霉菌酵母的生长繁殖,使得石榴籽粒保持较好的感官品质,延缓石榴籽粒的衰老进程。  相似文献   

Definition and Selection of Inferior Position Industries in old Industrial Basement     

CHEN Xun 《保鲜与加工》2002,(11):34-37

According to the industrial character of old industrial basement, such as Chongqing, we select preliminarily the candidated inferior position industries by using DEA analyse model. The synthetical basical coefficient of candidated inferior industries is obtained by analysing their indices such as income elasticity of demand, compared superiority coefficient, compared rate coefficient and technical progress rate, etc. Finally determine inferior industries which was supposed to exit by considing fully society progress and continuable development. A new idea are provided for industral structure adjustment for old industrial basement in China.  相似文献   

Empirical Research on Factors Influencing Stock Returns of Listed Companies in China     

LI Xun  CAO Guo-hua 《保鲜与加工》2006,(10):143-147153

  相似文献   

Dropwise Condensation Heat Transfer Coefficient on the Horizontal Circular Surface with Radial Gradient Surface Energy     

GU Yang-biao  LIAO Qiang  ZHU Xun  WANG Hong 《保鲜与加工》2006,(5):48-51

A theoretical model is developed for the dropwise condensation heat transfer on the horizontal circular surface with radial gradient surface energy based on the heat transfer model of individual condensate drop and the size distribution model of condensate drop on homogeneous condensation surface.The effect of variation of contact angle on the gradient surface on condensation heat transfer and condensate drop size distribution is taken account of in this model.The theoretical calculation method was obtained to predict the dropwise condensation heat transfer coefficient on the horizontal circular surface with radial gradient surface energy under various wall subcooled temperature,contact angle profile on wall surface,and working fluid.The effects of surface energy gradient,wall subcooled temperature,and thermophysical properties of condensate on the condensation heat transfer are discussed respectively.The calculation results show that the condensation heat transfer coefficient increases slightly with the increase of wall subcooled temperature.As latent heat and surface tension increasing,the condensation heat transfer coefficient increases.A larger surface energy gradient induces a larger condensation heat transfer coefficient.  相似文献   

Analysis of Social Appraisal Index System in Public Project     

HUA Yong-jun  CHEN Xun  ZHANG Jian 《保鲜与加工》2005,(7):145-147

The social evaluation theories of public project still have not been standardization today, and have effects on the process of the construction to city. One of the main reasons lays in that the index system, about the social evaluation theories of public project, connects with many fields, has many non-economic index and difficult in measuring the every index. This article is to mark off the main factors involved in social evaluation effectually, to set up index system of social evaluation practically, and to settle firm ground for methodology and theory of public project.  相似文献   

A Method of Analyzing the Parameter Sensitivity of Large Rotor Systems and Its Application     

Huang Wenzhen Huang Buyu Dong Xun 《保鲜与加工》1993,(1):72-77

Based upon the eigenvalue sensitivity theory of non-self-adjoit system and the component mode synthesis method,a new technique is developed to analyze the eigenvalue sensitivity to variation of parameters of large rotor-bearing system. As an application, the influence of the journal bearing parameters on the stability of a large domestic turbine-generator set is analyzed and discussed.  相似文献   

茶树钾营养特性的研究   总被引：5，自引：0，他引：5  

王晓萍  阮建云 《作物学报》1995,21(3):324-329

本文对我国主要茶区茶园土壤的含钾状况，茶树品种和土壤水分对茶树吸收钾的影响和茶园施肥钾肥的增产效果进行了系统研究。结果表明，我国有近６０％的茶园土壤处于罐钾状态，这已成为限制茶叶高产优质的主要障碍因子之一。不同品种茶树对钾的吸收具有明显差异，它们对钻的最大速率Ｖｍａｘ为龙井４３＞碧云＞苹云＞碧峰＞菊花春，米氏亲和常数Ｋｍ为碧峰＞碧云＞龙井４３＞苹云＞菊花春。干旱处理的茶树体内累积的钾和叶片的含钾量  相似文献