家蚕脂蛋白基因BmLp-c21的表达分析及蛋白质亚细胞定位

家蚕30 kD脂蛋白家族在家蚕的生长发育过程中具有重要的生理功能。从家蚕蛹cDNA文库中克隆到一条编码30kD蛋白质的基因cDNA序列,该序列全长2 803 bp,ORF为792 bp,编码含263个氨基酸残基的脂蛋白(low molecular 30K lipo-protein pBmHPC-21;GenBank登录号:Q00801),命名为BmLp-c21。将BmLp-c21克隆至原核表达载体pET-28a(+),转化E.coli Rosetta(DE3),IPTG诱导表达后通过亲和层析得到纯化的融合蛋白,并免疫新西兰大白兔制备多克隆抗体。亚细胞定位显示BmLp-c21主要存在于细胞质中,呈点状或者片状分布。通过实时荧光定量PCR和Western blotting方法,分别检测到家蚕不同发育时期和5龄幼虫不同组织中BmLp-c21 mRNA的转录水平与蛋白质的表达水平存在较大差异,在蛹期及5龄幼虫血淋巴中的mRNA转录水平和蛋白质表达水平最高。初步推测BmLp-c21在家蚕蛹的变态发育过程以及蚕体脂质的运输方面具有重要的功能。     

高山草原条件下苜蓿种质抗霜霉性评价

对引自国内外的94个苜蓿品种在海拔3000m的甘肃桑科摹试验站进行了高山草原条件下的霜霉病抗性评价。结果表明,品种之间的抗病性差异颇大,发病率为0～100%,严重度为1～4。通过对各品种的发病率和严重两项指标进行聚类分析,按照抗病性从高到低将94个品种分为免疫、高抗、中抗、抗病、感病、中感、高感和极感8个抗性类群;其中：阿尔古奎斯、巴瑞尔,阿毕卡、安古斯、日本1品种、伊鲁瑰斯、布韦兹、托尔、81-89美国、CP4350萨蓝纳斯、贝维、润布勒、兰热来恩德和威斯康辛等14个品种属免疫类群;普劳勒、班纳、L2-1079匈牙利和兴平4个品种属高抗类群;肇东、陇中和陇东3个品种属高感类群;78-27捷克、阿波罗、准格尔、陕北和河西5个品种属极感类群。6～8月份病害随着生长期的推移而呈逐渐加重的趋势。绝大多数极感品种和所有高感品种均为国内品种,而所有免疫品种和绝大多数高抗品种均为国外引进品种,这种由于品种的来源不同而引起的抗病性差异很可能与苜蓿霜霉菌的高度寄生专化性有关。     

Molecular genetics of behaviour: research strategies and perspectives for animal production

Genetic factors are undoubtedly involved in inter-individual variability of the behaviours that may be important for livestock production, as shown by pedigree studies, comparison of genetic stocks raised in the same environment, and selection experiments. The knowledge of gene polymorphisms responsible for genetic variability would increase the efficiency of selection, as shown for instance by the identification of the ryanodine receptor gene that harbours the mutations responsible for the porcine stress syndrome, that allows the eradication of the susceptibility allele. One strategy is to screen systematically the genes that are known to be involved in regulation of behaviour (functional candidate genes). This strategy is however very difficult for most behavioural traits, since behaviour is an emerging function from the whole brain/body and the molecular pathways involved in genetic variability are very poorly understood. Another strategy is to investigate linkage between trait variation and genetic markers in a segregating population (usually an intercross or backcross between two strains or breeds contrasting for the trait under study). It allows the detection of genomic regions influencing that trait (quantitative trait loci or QTL), and further investigation aims at the identification of the gene(s) located in each of these regions and the molecular polymorphisms involved in phenotypic variation. Although many QTL have been published for behavioural traits in experimental animals, very few examples are available where strong candidate genes have been identified. Further progress will be very much dependent upon the careful definition of behavioural traits to be studied (including their importance for animal production), on the reliability of their measurement in a large number of animals and on the efficient mastering of environmental factors of variability. The fast increase in the knowledge of genome sequence in several species will undoubtedly facilitate the application to farm animal species of the knowledge obtained in model organisms, as well as the use of model organisms to explore candidate genes detected by QTL studies in farm animals.     

鸡大肠杆菌O78对喹诺酮类药物高耐药株的分子鉴定

就临床分离的鸡大肠杆菌O78对喹诺酮类药物的最低抑菌浓度（MIC）进行了测定，得到对喹诺酮类药物有不同耐药水平的细菌23株。根据GenBank已公布的QRDRs序列，设计了分剐扩增gyrA、gyrB、parC和parE基因的4对引物，以筛选的23株耐药菌DNA为模板，进行了PCR扩增。序列分析及AcrA的Western blotting检测结果表明，临床分离的鸡大肠杆菌对喹诺酮类药物的耐药水平与GyrA和ParC的突变密切相关，而AcrAB外输泵的表达水平无显著变化。提示临床分离的鸡大肠杆菌O78的耐药水平与喹诺酮类药物的选择性压力有关，它诱导了DNA旋转酶和拓扑异构酶IV的基因突变，可能不能激活AcrAB外输泵。     

新疆不同地区牛源大肠杆菌耐药性分析

This study was aimed to investigate the resistance of Escherichia coli isolates from cattle farms in Xinjiang to antibiotics. The resistances of Escherichia coli isolates from cattle farms in five regions to commonly used antibiotic were determined using microdilution broth assay. Escherichia coli isolates were highly resistant to ampicillin (up to 39.3%) in all regions, and existed multidrug-resistant, but zero resistance was the dominant. Escherichia coli isolates from E cattle farm in Cherqi town, Baicheng county, exhibited resistant to three kinds of quinolones (6.7%). The results suggested that the resistances of Escherichia coli isolates to commonly used antibiotics were not serious in Xinjiang.     

H3亚型猪流感病毒荧光定量PCR检测方法的建立

通过RT-PCR方法克隆了H3亚型猪流感病毒HA基因一段靶序列,构建重组质粒作为标准阳性模板.根据GenBank中的H3亚型猪流感病毒HA基因保守序列设计了用于FQ-PCR的1对引物和1条TaqMan探针.通过条件优化,以10倍系列稀释的质粒为标准品进行荧光定量PCR扩增,并制作标准曲线,建立了检测H3亚型猪流感的荧光定量PCR方法.结果表明,该方法检测灵敏度可达1.0×100拷贝/μL,线性范围为109～100,达10个数量级;对起始浓度为1.0×109、1.0×108、1.0×107拷贝/μL的标准品的最终实际测得值(Ct)分别为13.68,18.21和20.57;变异系数分别为0.31%、0.17%和0.12%,均小于5%,说明此方法具有良好的准确性和重现性.对阳性组织病料的检测表明,该方法的检测灵敏度高出常规PCR,与套式PCR具有相近的灵敏度.     

Non-iterative variance component estimation in QTL analysis

In variance component quantitative trait loci (QTL) analysis, a mixed model is used to detect the most likely chromosome position of a QTL. The putative QTL is included as a random effect and a method is needed to estimate the QTL variance. The standard estimation method used is an iterative method based on the restricted maximum likelihood (REML). In this paper, we present a novel non-iterative variance component estimation method. This method is based on Henderson's method 3, but relaxes the condition of unbiasedness. Two similar estimators were compared, which were developed from two different partitions of the sum of squares in Henderson's method 3. The approach was compared with REML on data from a European wild boar × domestic pig intercross. A meat quality trait was studied on chromosome 6 where a functional gene was known to be located. Both partitions resulted in estimated QTL variances close to the REML estimates. From the non-iterative estimates, we could also compute good approximations of the likelihood ratio curve on the studied chromosome.     

毒死蜱对桔小实蝇体内代谢酶的影响

为了解毒死蜱对桔小实蝇体内代谢酶的影响,采用不同浓度的毒死蜱对桔小实蝇敏感种群进行连续处理,测定其体内的羧酸酯酶（CarE）、谷胱甘肽S-转移酶（GSTs）、过氧化氢酶（CAT）、过氧化物酶（POD）的活性。结果表明：随着毒死蜱浓度的增加,桔小实蝇的敏感性出现逐渐下降的趋势,体内的CarE活性得到显著提升（F=43.21,p=0.024）,抗性倍数为1.74,GSTs活性也显著上升（F=47.73,p=0.018）,抗性倍数为1.98,但对CAT、POD的提升作用较小,表明毒死蜱对桔小实蝇体内的CarE、GSTs活力具有一定的诱导作用。     

细菌耐药质粒消除方法研究进展

常见的病原微生物多重耐药性的增加以及多重耐药菌株的出现,给动物养殖业造成了巨大的经济损失。细菌的耐药性与其自身携带的耐药质粒关系密切。论文从细菌耐药现状,细菌的耐药性和耐药质粒的关系,耐药质粒的物理、化学、中草药消除方法这几个方面进行综述,旨在寻求一种安全有效的消除方法消除耐药质粒,为今后相关的研究提供参考。