PCR方法鉴别肉骨粉中的动物成份种类

为防范疯牛病传入我国,有必要鉴别肉骨粉中动物成份的种类。针对牛、羊、猪和鸡四种动物的特异性核苷酸序列,分别选用和设计了四对特异性引物,用试剂盒提取四种动物的肉骨粉或者处理过的肉组织中的DNA,然后进行PCR扩增,所扩增的目的片断大小分别为271bp、199bp、196bp、148bp,运用PCR技术建立了鉴定这四种动物源性成分的方法。结果分别扩增出四种动物的特异性条带,证明该PCR方法具有很高的特异性和敏感性,而且成本低廉,简便易行,因此可以作为鉴别肉骨粉种类的常规方法。     

Effect of ruminal dosing of mechanical stimulating brush on rumination time, ruminal passage rate and rumen fermentation status in Holstein steers fed a concentrate diet

This experiment was conducted to determine the effect of ruminal dosing of a mechanical stimulating brush on rumination time, ruminal passage rate and rumen fermentation status in steers fed a concentrate diet at maintenance level. Animals were dosed three Rumen Faibu (RF) per head through the rumen fistulae (RF treatment) and not dosed (control) in a change‐over design. The organic cell wall content of the concentrate diet was 12.7% of dry matter. Daily time spent on rumination was very short in both treatments with 24 min in RF treatment and 15 min in control. The turnover rate of ruminal fluid in RF treatment was higher than that in control. There were no differences in ruminal pH and total volatile fatty acid concentration between RF treatment and control. Acetic and butyric acid concentrations were not different between the treatments. Propionic acid concentration tended to be higher in the animals on RF treatment than in control animals. The RF dosing in Holstein steers fed a low fiber diet did not affect the rumination time, but increased rumen digesta passage rate and ruminal propionic acid production.     

甘肃省水资源时间序列模型及其变化规律研究

水文水资源序列是一个具有周期变化、随机变化和递增或递减趋势变化的复杂时间序列。作为一种近似,可以把水文序列{Qt}分为趋势序列{Nt}、周期序列{Pt}和随机性的平稳时间序列{St},即式:Qt=Nt+Pt+St。采用时间序列分析的方法,建立了甘肃省水资源的时间序列模型,分析了其变化规律,结果表明,甘肃省水资源呈明显下降趋势,并存在一定的周期变化规律。     

迪卡配套系猪RYR1的PCR分析及其利用的研究

试验从鸡东县永安猪场随机抽取迪卡配套系猪B系和E系42头，通过PCR扩增技术和酶切鉴定．采用克隆技术进行测序，来检测是否含有氟烷基因序列。经过以上技术检测这42头猪均没有氟烷基因。     

猪肺炎支原体黏附因子基因R1R2区的克隆及表达

根据GenBank登录的猪肺炎支原体232株P97基因和J株黏附因子基因设计了1对引物，以我国猪肺炎支原体Z株(强毒株)基因组DNA为模板，通过PCR方法扩增了该株黏附因子基因的部分序列。经序列分析后，重新设计了1对带有EcoRI和HindⅢ酶切位点的引物，并经引物的定点突变，PCR扩增了Z株黏附因子的R1R2区。扩增产物经双酶切后克隆到表达载体pET-32(a) 中。该重组质粒经酶切鉴定后，将其具有正确阅读框架的重组质粒转化到大肠杆菌BL21(DE3)感受态细胞，37℃下经IPTG诱导表达，得到相对分子质量约29000的融合蛋白，表达量约为11％。     

小鹅瘟PCR诊断方法的建立和初步应用

根据发表的鹅细小病毒B株VP3基因序列 ,设计合成了 1对寡聚核苷酸引物。以国内分离的小鹅瘟病毒为摸板 ,筛选了 (PCR)最佳反应条件 ,并进行了敏感性、特异性和初步应用试验。结果表明 ,在 4 0 μL体积中 ,PCR最佳系统组成为 2uTaq酶、0 5mol/LdNTP和 2 0 pmol引物 ;最佳反应参数为 96℃变性 4 0s ,5 4℃退火 1min ,72℃延伸 1min ,30个循环。本方法可检出 2 5× 10 -1 5EID50 小鹅瘟病毒 ,并且仅能从小鹅瘟病毒的鹅胚培养物中扩增出与设计值大小相同的 375bp核苷酸片断 ,对照病毒扩增结果为阴性。通过对 6份临床病料的检查 ,2份呈阳性 ,其中 1份来自有典型小鹅瘟症状的雏鹅 ,1份来自无典型小鹅瘟症状的雏鹅。由此说明 ,本试验所建立的诊断小鹅瘟的PCR方法具有潜在的临床应用价值     

单抗酶联试剂盒与PCR方法快速检测沙门氏菌的比较

本文对单抗酶联试剂盒与PCR方法快速检测沙门氏菌进行了比较。将沙门氏菌与大肠杆菌以不同比例混合后 ,分别用ELISA和PCR法检测 ,在 1∶1 0 0的比例时 ,用这 2种方法均能检测出阳性样品 ;在 1∶1 0 0 0的比例时 ,用ELISA法检测为阴性 ,而用PCR法则能检测出。检测鲜牛奶、龙虾、虾仁、熟食制品等样品 ,2种方法的符合率达 97 6 %。对冻虾仁、人粪便样品的前增菌、选择性增菌、后增菌过程进行同步检测 ,在样品的前增菌液 ,直接ELISA法检测的OD值小于 0 5 ,而PCR法扩增能出现特异条带 ,经国标方法验证为阳性 ,证实PCR法的敏感性优于直接ELISA法 ,而在选择性增菌液和M 肉汤后增菌液中 ,2种方法均能检出阳性样品。     

JEV、PPV、PRRSV、PRV多联PCR的应用研究

用 JEV、PRRSV二联 PCR,PPV、PRV二联 PCR以及这 4种病毒 4联 PCR对来自内蒙、广州、广西、天津、北京、吉林病料进行检测 ,共检了 1 4 6份病料。其中 PRRSV阳性 7份 ,PPV阳性 1 2份 ,PRV阳性 2 1份 ,PRV和 PPV混合感染 4份。并对 5份人工接种 JEV小鼠病料检测 ,其中 4份为阳性。随后对部分阳性 PCR扩增产物进行点杂交和核苷酸测序鉴定 ,证实了 PCR扩增准确性。对内蒙 PRRSV阳性扩增带测序结果显示 ,我国流行 PRRSV为美洲型 ,在扩增片段的核苷酸序列上有 3个碱基差异     

实广义正定矩阵的若干性质

矩阵∈AR~(n×n)称为实广义正定矩阵,如果对任意非零向量 X∈R~n,有XTAX>0成立。本文讨论了矩阵的kronecker积。Hadamard积和矩阵乘积的正定性,给出相应一些性质。     

Trends in NE Atlantic landings (southern Portugal): identifying the relative importance of fisheries and environmental variables

Time series of commercial landings from the Algarve (southern Portugal) from 1982 to 1999 were analyzed using min/max autocorrelation factor analysis (MAFA) and dynamic factor analysis (DFA). These techniques were used to identify trends and explore the relationships between the response variables (annual landings of 12 species) and explanatory variables [sea surface temperature, rainfall, an upwelling index, Guadiana river (south‐east Portugal) flow, the North Atlantic oscillation, the number of licensed fishing vessels and the number of commercial fishermen]. Landings were more highly correlated with non‐lagged environmental variables and in particular with Guadiana river flow. Both techniques gave coherent results, with the most important trend being a steady decline over time. A DFA model with two explanatory variables (Guadiana river flow and number of fishermen) and three common trends (smoothing functions over time) gave good fits to 10 of the 12 species. Results of other models indicated that river flow is the more important explanatory variable in this model. Changes in the mean flow and discharge regime of the Guadiana river resulting from the construction of the Alqueva dam, completed in 2002, are therefore likely to have a significant and deleterious impact on Algarve fisheries landings.