猪肺炎支原体膜蛋白的电泳分析及免疫印迹检测

以猪肺炎支原体Z株为试验材料,采用SDS-PAGE和Western印迹对该菌膜蛋白进行了分析。研究表明:猪肺炎支原体经A_(26)液体培养基培养,15000 r/min离心收集菌体细胞,低渗超声法破膜获得膜制剂后,应用12.5％的凝胶对膜蛋白进行SDS-PAGE分离,在电泳图谱上呈现出36条蛋白带,相对分子质量范围为1.18×10~4～9.12×10~4。同时以膜制剂作为抗原,分4次免疫试验兔,心脏采血,提取抗血清,并分离纯化得到IgG,然后进行Western印迹法检测。其结果发现在SDS-PAGE分离得到的36种膜蛋白(或多肽)中,有6种蛋白具有免疫原性。     

山羊TNNT3基因可变剪切及其对骨骼肌细胞分化的作用

【目的】快速骨骼肌肌钙蛋白T(fast skeletal troponin T3,TNNT3)作为肌钙蛋白(troponin, Tn)家族成员,调节横纹肌收缩、参与骨骼肌的生长发育并影响家畜肉质性状。通过获得山羊TNNT3基因的可变剪切体,分析山羊TNNT3基因可变剪切的表达模式及其在肌细胞分化中的作用,深入解析TNNT3基因在山羊骨骼肌生长发育过程中的作用机制。【方法】基于NCBI已公布山羊TNNT3基因(NM₀01314210.1)和牛TNNT3基因(XM₀10821200)mRNA序列,使用软件Primer Premier6.0设计引物,以简州大耳羊胚胎期和出生后7个阶段骨骼肌为试验材料,克隆测序获得山羊TNNT3基因的CDS区可变剪切体,利用软件ORF Finder、EditSeq、DNAMAN、ClustalW和MEGA_X10.1.8等对序列进行生物信息学分析;进一步设计实时荧光定量(real-time PCR,RT-qPCR)及半定量引物,研究TNNT3基因剪切体在7个不同组织(背最长肌...     

民族地区培育村镇银行的法律问题研究

为了克服制约村镇银行发展的诸多不利因素,从法律角度提出了在民族地区培育村镇银行的对策：一是完善村镇银行立法;二是放宽村镇银行的发起人资格限制;三是完善财政支持制度;四是依据税法给予民族地区村镇银行税收优惠;五是探索建立存款保险制度;六是重构村镇银行监管法律责任制度;七是健全政策性农业保险制度,构建农业巨灾风险分散机制;八是对民族地区村镇银行实施倾斜性的货币政策;九是制定和完善相关法律法规和规章。     

西部12省市农村居民生活消费支出现状的比较研究

选取西部12省市的农村居民和各地区农村居民家庭平均每人生活消费支出的数据,采用面板数据模型分析居民收入结构对生活消费支出的影响实证研究。结果表明：农村居民的工资性收入、家庭经营纯收入、财产性收入和转移性收入对生活消费支出都有显著影响,且财产性收入和转移性收入的边际消费倾向都大于1。     

武陵山民族地区新型城镇化问题调研报告

新型城镇化将是我国今后几十年经济增长的巨大引擎,也是促进民族地区快速发展的历史机遇.通过对武陵山民族地区6个县新型城镇化的实地调研,了解到改革开放以后这个地区的城镇化建设有了长足进展,当地政府对推进新型城镇化建设的积极性空前高涨.但面对“四化”的新形势,武陵山民族地区新型城镇化建设的制约因素十分明显,主要是城镇规划无序滞后,产业发展先天不足,民族文化传承任重道远,建设资金欲求无门,政策“洼地”严重失衡.根据以上情况,提出了推进民族地区新型城镇化建设建议.     

Generation of mouse monoclonal antibodies specific to tilapia immunoglobulin using fish immunoglobulin/BSA complex for monitoring of the immune response in Nile tilapia Oreochromis niloticus

The simple immunoprecipitation method was used to isolate tilapia immunoglobulin (Ig) for immunization in order to produce monoclonal antibodies (MAbs) specific to tilapia Ig. First, the tilapia antiserum against bovine serum albumin (BSA) was prepared by peritoneal injection of BSA into tilapia, and the tilapia anti‐BSA antiserum was used to precipitate BSA to form the Ig/BSA immune complex. The Ig/BSA immune complex was then injected into Swiss mice for hybridoma production. After fusion, three hybridoma clones producing MAbs specific to the tilapia antibody were selected by dot blot and Western blot. All MAbs (101A, 59G, and 11A) were bound specifically to the heavy chain of immunoglobulin M (IgM). The MAbs 101A and 59G demonstrated twofold higher affinity than MAb 11A and the commercialized antibody. However, MAbs 11A could also bind to the heavy chain of IgM in Asian seabass, Lates calcarifer, as well. These MAbs can be used to monitor the immune responses of individual fish by indirect ELISA upon exposure to various antigens.     

盛夏西太平洋副热带高压对陕西降水的影响

利用1961-2008年陕西78个气象站7-8月降水资料、西太平洋副高逐月指数资料及500hPa位势高度场和700hPa风场月平均再分析资料,通过相关、回归分析、小波凝聚谱分析等方法探讨盛夏西太平洋副热带高压对陕西降水的影响及机制,为盛夏陕西降水的气候预测提供依据.结果表明,盛夏副高强度、脊线、西伸脊点指数与陕西降水关系密切,副高偏强偏西时陕西降水偏多,副高偏北时陕西北部降水偏多、南部偏少.8月副高3个指数与陕西降水的相关关系更为密切,通过显著性检验的站点明显增多.副高的强度和位置变化引起副热带和中高纬度大气环流的异常,进而影响陕西降水.8月副高偏强偏西时,巴尔喀什湖阻高增强,700hPa西南季风的水汽输送增强,陕西位于冷暖空气交汇区,降水偏多;副高偏南时,水汽辐合区位于陕南,陕西南部多雨、北部少雨;副高偏北时,水汽辐合区位于陕北,陕西北部多雨、南部少雨.8月副高强度指数与同月陕西降水的显著共振周期为7 ～8a,且副高强度变化超前于陕西降水变化1.26a,说明8月副高强度的年际变化是引起同月陕西降水年际变化的重要原因.     

Field compaction at different soil-water status: effects on pore size distribution and soil water characteristics of a Rhodic Ferralsol in Western Cuba

The level of compaction induced on cultivated fields through trafficking is strongly influenced by the prevailing soil-water status and, depending on the attendant soil degradation, vital soil hydraulic processes could be affected. Therefore, understanding the relationship between field soil-water status and the corresponding level of induced compaction for a given load is considered an imperative step toward a better control of the occurrence of traffic-induced field soil compaction. Pore size distribution, a fundamental and highly degradable soil property, was measured in a Rhodic Ferralsol, the most productive and extensively distributed soil in Western Cuba, to study the effects of three levels of soil compaction on soil water characteristic parameters. Soil bulk density and cone penetration index were used to measure compaction levels established by seven passes of a 10 Mg tractor at three soil-water statuses corresponding to the plastic (Fs), friable (Fc) and relatively dry soil (Ds) consistency states. Pore size distribution calculated from soil water characteristic curves was classified into three pore size categories on the basis of their hydraulic functioning: >50 μm (f_>50 μm), 50–0.5 μm (f_{50–0.5 μm}) and <0.5 μm (f_<0.5 μm). The greatest compaction levels were attained in the Fs and Fc soil water treatments, and a significant contribution to compaction was attributed to the existing soil water states under which the soil compaction was accomplished. Average cone index (CI) values in the range of 2.93–3.70 MPa reflected the accumulation of f_<0.5 μm pores, and incurred severe reductions in the volume of f_>50 μm pores in the Fs and Fc treatments, while an average CI value of 1.69 MPa indicated increments in the volume of f_{50–0.5 μm} in the Ds treatment. Despite the differential effects of soil compaction on the distribution of the different pore size categories, soil total porosity (f_Total) was not effective in reflecting treatment effects. Soil water desorption at the soil water potentials evaluated (0.0 to −15,000 cm H₂O) was adversely affected in the f_<0.5 μm dominated treatments; strong soil water retention was observed with the predominance of f_<0.5 μm, as was confirmed by the high water content at plant wilting point. Based on these findings, the use of field capacity water content as the upper limit of plant available soil water was therefore considered inappropriate for compacted soils.     

Molecular cloning and expression of a novel eukaryotes elongation factor1A gene (ZjeEF-1α) from Chinese jujube in response to phytoplasma infection

Eukaryotic translation elongation factor 1A (eEF1A) in plant is a multifunctional protein. In our previous study, we identified a 651-bp EST sequence of eEF-1α from a suppression subtractive hybridization (SSH) library of Chinese jujube (Ziziphus jujuba Mill.), which was established under jujube witches' broom (JWB) phytoplasma stress. Here we isolated the eEF1A gene (ZjeEF-1α, JF488063) from Chinese jujube by single oligonucleotide nested (SON)-PCR and rapid amplification of cDNA ends (RACE). ZjeEF-1α contains an open reading frame (ORF) of 1344 bp, encoding a polypeptide of 447 amino acids with an estimated molecular mass of 49.29 kDa and a theoretical pI of 9.10. ZjeEF-1α shared over 92% similarity with eEF-1α proteins from other 24 eukaryotes including plants and animals. Search Pfam predicted that ZjeEF-1A contains three typical eEF1A functional domains. Moreover, the corresponding genomic sequence of ZjeEF-1α was also sequenced, in which a 115-bp intervening sequence with splicing site GT-AG was found. Real-time quantitative PCR (RT-qPCR) and western blotting assays revealed that the expression of ZjeEF-1α was enhanced in resistant cultivar ‘Xingguang’ and decreased in sensitive cultivar ‘Zanhuangdazao’ at early stages of phytoplasma infection. It is proved that ZjeEF-1A expression is early response to phytoplasma infection and its possible functions in resistant process to JWB phytoplasma are discussed.     

犬瘟热病毒H基因的序列分析与表达

以临床疑似犬瘟热病犬外周血总RNA为模板,采用RT-PCR方法扩增犬瘟热病毒(CDV)血凝素(H)基因。序列测定和分析表明该病料犬瘟热病毒H基因序列与国内其他地区分离到的毒株同源性较高(94.6%~99.2%),而与Onderstepoort株、Convac株等疫苗毒株的同源性较低(90.4%~91.2%)。分子遗传进化分析显示所有毒株可分为7个大的分支,且各分支间具有一定的地域性,其中待检病料中CDV与大多数中国分离株一样处于Ⅰ型。进一步以H全基因为模板,截短表达其3′端816 bp、459 bp两个片段,并克隆入原核表达载体pET30 a进行表达。结果显示它们分别能表达大小约为36.4 ku和22.2 ku的融合蛋白。免疫转印表明该纯化蛋白均可与犬瘟热病毒抗血清发生阳性反应,说明重组蛋白具备抗原性,可用于进一步的血清学研究。