PRRSV重组N蛋白表达、纯化及间接ELISA检测试剂盒的研制

采用RT-PCR扩增出大小409bp猪繁殖与呼吸综合征病毒核衣壳蛋白基因（PRRSV N gene）,将该基因克隆到表达载体pGEX-KG,构建重组表达载体pGEX-KG-N,转化表达菌株BL21（DM3）诱导表达,经SDS-PAGE和Western blot分析表明,重组N蛋白获得了高效表达,融合蛋白相对分子质量为41 000,并具有良好的免疫学活性。扩大诱导培养,收集菌体,提取包涵体,用GST-Protein Purtification Kit亲和层析纯化目的蛋白,SDS-PAGE电泳检测纯度达到90%以上,可满足诊断抗原质量要求。用纯化PRRSV重组核衣壳蛋白GST-N为包被抗原,建立检测PRRSV血清的间接ELISA诊断方法,并组装ELISA试剂盒。优化后抗原最适包被质量浓度为2.5mg/L;血清最适稀释度为1∶100;对血清样本检测临界值为0.224;与美国IDEXX公司的HerdChek ELISA试剂盒符合率为92%,特异性为95%,敏感性为90%;与猪细小病毒、猪伪狂犬病病毒、猪乙型脑炎病毒、猪圆环病毒、猪瘟病毒等常见猪繁殖障碍病标准阳性血清无交叉反应。ELISA试剂盒批内和批间变异系数分别为3.44%～6.34%和5.04%～7.64%。置4℃条件保存12个月,试剂盒稳定性无明显改变。该试剂盒对350份临床疑似血清检出率为88.6%。研制的ELISA试剂盒为PRRSV临床血清抗体检测及流行病学调查提供了技术手段。     

抵抗素样Relmβ分子研究进展

Relmβ亦称发现于炎症带2(found in inflammatory zone2,FIZZ2),是一种富含半胱氨酸的分泌蛋白。目前的研究表明,Relmβ在许多方面发挥着非常重要的作用,包括胰岛素抵抗,抵制肠腔寄生线虫的感染,预测胃肠道肿瘤的发生,促进小鼠结肠炎和回肠炎模型中的炎症反应,诱导肺部纤维化、缺氧肺血管的重建,促进气道重塑等。通过对Relmβ分子的结构、表达分布、生物学功能的研究进展进行归纳,为Relmβ分子在生物学和医学方面更深入的研究提供参考。     

复杂盐碱生态条件的人工模拟及其对羊草生长的影响

将ＮａＣｌ、ＮａＨＣＯ３、Ｎａ２ＳＯ４和ＮａＣｏ３按不同比例混和,模拟出３０种盐度和ＰＨ各不相同的盐碱生态条件,并用以处理并草苗。测定羊草苗的存活率、分蘖数、根戏数及相对生长率等生长指标;分析盐度和ＰＨ对羊草生长失作用及其盯关性。结果表明：３０处处理均匀覆盖了总盐度５０－３５０ｍｍｏｌ／ＬＰＨ７．１４－１０．８１范围内的各处盐碱条件,羊草的各项生长指标均随盐浓度及Ｐ增高而下降;单纯高ＰＨ域盐度对羊     

不同季节收获的星星草对绵羊采食量和养分消化率的影响

旨在研究不同季节收获的星星草(Puccinellia tenuiflora)对绵羊采食量及养分消化率的影响。选用平均体重为30.37±2.15kg的乌珠穆沁公羊12只,随机分为4个处理,每个处理3个重复,分别饲喂四季收获的星星草。试验绵羊采取单笼饲养,预饲期15d,正式试验期7d,自由采食和饮水,采用全收粪法进行消化代谢试验。结果表明:绵羊四季采食星星草干物质含量分别为412.42、639.23、741.49和321.30g/d,秋季显著高于春季和冬季的干物质采食量(P<0.05);秋季星星草有机物消化率显著低于其他季节(P<0.05);不同季节的星星草对绵羊的碳、氮采食量均有显著影响(P<0.05),饲喂不同季节星星草的绵羊均处于氮的负平衡状态,分别为-1.12、-0.78、-0.13和-3.16。总之,星星草低的含氮量和采食量是限制星星草饲喂价值的重要因素。     

犬黄嘌呤尿结石症的诊断:36例临床病理报告

36只尿石症病犬表现体温下降,剧烈呕吐,食欲废绝,血清尿酸、肌酐、尿素氮升高,尿沉渣中有众多结晶体析出,经尿液和血清生化指标测定、B超探察、X线影像并结合饮食结构调查,确诊为黄嘌呤结石症。     

细胞壁降解相关酶以及在反刍动物应用中活性测定标准化的建议

针对当前国内外饲料酶制剂在反刍动物应用上结果不一致的现象,本文从酶制剂的角度,简要地分析了可能引起这些不一致的原因,同时就此总结了与细胞壁降解相关的主要的酶及其功能,并给出其活性测定标准化建议,为酶制剂的开发和应用提供参考。     

Cloning of Buffalo AQP9 Gene and Analysing its Expression in the Buffalo Tissues

Cloning buffalo AQP9 gene and analyzing its expression in buffalo tissues.A pair of primers was designed according to the released bovine AQP9 sequences in GenBank,which was used to clone buffalo AQP9 gene.The AQP9 gene was amplified by RT-PCR,whose nucleotide sequence and protein structure were analyzed by bioinformatics methods.The expression of AQP9 in buffalo tissues was assayed by Real-time quantitative PCR.The expression of AQP9 gene in buffalo ovary and testis tissue was detected by immunohistochemical staining method.The results showed that the cloned ORF length of buffalo AQP9 gene was 888 bp,which coded 295 amino acids.The results of multiple sequence comparison showed that the nucleotide sequence of buffalo AQP9 shared 99%,90%,97% and 88% homologeous compared with that of Bos taurus,Sus scrofa,Ovis ariessis and Homo sapiens,respectively,while shared 99%,86%,97%,83% homologeous for amino acids,respectively.Phylogenetic tree analysis indicated that AQP9 gene was highly conservative in the evolutionary process.Real-time quantitative PCR results showed that AQP9 gene expressed in buffalo liver,lung,brain,skin,testis and ovary tissues with different levels,had the most abundant expression in liver,followed by in skin and testis,less observed in lung and ovary.The results of immunohistochemical staining showed that the expression of AQP9 protein varied with the development of buffalo ovarian tissue,and gradually enhanced with follicle development.In testicular tissue,AQP9 protein expressed in spermatocyte and leydig cells of developmental stage testis.These results indicated that we had successfully cloned buffalo AQP9 gene sequences.The expression and its function of AQP9 in buffalo ovaries and testes might play an important role in follicle development and spermatogenesis.     

鹅副黏病毒与新城疫病毒对鸡胚和鹅胚成纤维细胞的感染性分析

用鸡新城疫病毒F48E9,Komarov, LaSota,V4/66株和鹅副黏病毒SF02株分 别感染鸡胚和鹅胚成纤维细胞,用MTT显 色方法检测存活细胞数目。结果表明,不同 毒力新城疫病毒致细胞病变的作用不一样, 鸡新城疫强毒F48E9株和鹅副黏病毒SF02 株均引起两种细胞几乎全部死亡,感染动物 时,SF02与NDV强毒株的致病性有显著的 区别,但感染细胞时,二者并无明显差别; NDV与GPMV在感染水禽时的致病性差异 并不是因为诸如细胞膜受体等细胞水平的因 素不同造成的,可能与干扰素途径有关。     

命案攻坚战警犬显神威

北京市公安局刑侦总队警犬侦查训练大队认真贯彻落实“全国侦破命案工作会议”精神,从抓警犬“出现场率”入手,充分发挥警犬技术在侦破命案专项斗争中的作用,并取得显著成效。