转基因大豆低密度基因芯片的检测方法研究

目的建立Roundup Ready转基因大豆（简称RRS）的基因芯片检测方法,探讨其在转基因大豆检测中的应用。方法根据大豆中所转入的外源基因,选择camv35s启动子、nos终止子和cp4epsps基因,以大豆内源基因lectin基因为内参照基因设计了引物和探针,并制备了核苷酸芯片;通过多重PCR对样品核酸进行扩增和荧光标记后,将PCR产物与芯片杂交,检测大豆样品中所含的外源基因,并评价方法的灵敏度。结果检测转基因含量不同的RRS标准参考物,结果显示：camv35s启动子、nos终止子、外源基因cp4epsps检测灵敏度均可达0．45％。结论本研究所建立的基因芯片检测方法有良好的灵敏度及可重复性,有助于实现转基因大豆的高通量、高灵敏的检测。     

大豆深加工产品中外源基因检测灵敏度的研究

为了提高大豆深加工食品中外源基因的检测灵敏度.改良了普通PCR反应的引物,并设计了NEST-PCR反应程序.模拟食品加工中高温、高压等处理方法,将转基因大豆研磨成粉状,在高压灭菌锅内进行高温、高压处理.应用新设计的引物35SCP142、CPNOSl65进行外源基因检测,通过对引物的调整,检测灵敏度得到了明显的提高,可以检测到的转基因大豆含量仅为1%.且为经过12l℃,30min处理的样品中的外源基因.应用NEST-PCR的高灵敏度的特性.设计了NEST-PCR的两对引物35SCP318和35SCP114.并检测到外源基因的片段.将普通PCR的检测灵敏度和荧光定量PCR的灵敏度作比较,若引物设计合理,且PCR条件适合,普通PCR的检测灵敏度可以和荧光定量PCR的相媲美.     

大豆过氧化物酶基因的克隆及其表达载体的构建

大豆过氧化物酶(soybean peroxidase,sbp)基因的克隆及其表达载体的构建将有利于人们更深入的从分子生物学角度研究sbp 基因的结构与功能。首先从大豆根中获取总RNA,通过RT-PCR 获得sbp 基因。再将sbp 基因与表达载体pPICZα-A 双酶切后连接,构建重组表达载体pPICZα-A-sbp。将pPICZα-A-sbp 转化大肠杆菌筛选阳性克隆体并测序。结果表明：获得的sbp 基因序列与已报道的sbp[U51191(GmEpa1)]基因序列有92% 的同源性。重组表达载体pPICZα-A-sbp 的成功构建为其在毕赤酵母中表达奠定了基础。     

Primers for Amplifying an Alanine Racemase Gene Fragment to Detect E. coli Strains in Foods

Specific oligonucleotide primers for detecting Escherichia coli in various foods were designed based upon the conserved sequences of the E. coli air gene from positions 322 to 345 and from 664 to 687. Bacteria and food samples were treated at 100°C for 10 min in 1% Tween 20 containing 5% NaCl and 1 mM EDTA, then used as templates for polymerase chain reaction (PCR). The oligonucleotide primers were specific to E. coli, except for Shigella species, when tested with 67 strains of E. coli, including such serotypes as O157:H7 and O111, and 32 strains of non-E. coli species. The oligonucleotide primers could prove useful for detecting E. coli in beef, chicken, pork, tomato, soybean, potato, cow's milk, and egg.     

大豆油氢化反应釜数值模拟及工艺优化

以一级大豆油为液相、Pt/C催化剂为固相,釜体为圆柱体,釜体高度为180 mm,内径为120 mm,液面高度为130 mm,利用FIUENT软件对大豆油氢化反应釜进行液固两相数值模拟,发现倾斜式搅拌桨距反应釜底部高度80 mm、桨叶直径40 mm、搅拌速率300 r/min时流体流动及催化剂分布最佳,并以模拟的主要参数制备了高压反应釜。高压反应釜内一级大豆油添加量90.0 g、Pt/C催化剂添加量0.15%(m/m),充入8 MPa的CO2气体,后充入H2保持反应釜内总压为12 MPa,通过优化得出最佳反应温度97℃、反应时间87 min、搅拌速率285 r/min时,氢化后大豆油的碘值为79.50 g I2/100 g,说明模拟准确,为展示大型设备油脂氢化过程提供理论依据。     

应用通用引物PCR法检测葡萄球菌A、D和E型肠毒素基因

本实验采用20bp通用引物P3和P4的PCR方法对产A、D和E肠毒素的葡萄球菌进行了检测。结果表明,产SEE菌株能产生666bp的特异扩增片段,产SEA菌株产生666bp和约400bp两条扩增带,产SED菌株产生400bp片段;SEE菌株的扩增产物经EcoRV酶切能产生251和415bp两个片段。扩增敏感性实验表明,该方法可检出10~6个细菌。     

Accuracy of direct genomic values for functional traits in Brown Swiss cattle

In this study, direct genomic values for the functional traits general temperament, milking temperament, aggressiveness, rank order in herd, milking speed, udder depth, position of labia, and days to first heat in Brown Swiss dairy cattle were estimated based on ~777,000 (777K) single nucleotide polymorphism (SNP) information from 1,126 animals. Accuracy of direct genomic values was assessed by a 5-fold cross-validation with 10 replicates. Correlations between deregressed proofs and direct genomic values were 0.63 for general temperament, 0.73 for milking temperament, 0.69 for aggressiveness, 0.65 for rank order in herd, 0.69 for milking speed, 0.71 for udder depth, 0.66 for position of labia, and 0.74 for days to first heat. Using the information of ~54,000 (54K) SNP led to only marginal deviations in the observed accuracy. Trying to predict the 20% youngest bulls led to correlations of 0.55, 0.77, 0.73, 0.55, 0.64, 0.59, 0.67, and 0.77, respectively, for the traits listed above. Using a novel method to estimate the accuracy of a direct genomic value (defined as correlation between direct genomic value and true breeding value and accounting for the correlation between direct genomic values and conventional breeding values) revealed accuracies of 0.37, 0.20, 0.19, 0.27, 0.48, 0.45, 0.36, and 0.12, respectively, for the traits listed above. These values are much smaller but probably also more realistic than accuracies based on correlations, given the heritabilities and samples sizes in this study. Annotation of the largest estimated SNP effects revealed 2 candidate genes affecting the traits general temperament and days to first heat.     

用近红外仪快速测定油料及粕中蛋白质,脂肪和水分方法的探讨

利用近红外仪测定油料及粕中蛋白质、脂肪和水分，同常规实验室分析法对照比较，得到较为一致的结果，该方法可以替代常规实验室方法。     

可视芯片检测大豆、水稻和玉米中的转基因成分

本研究选取9种常见转基因食品外源基因,设计了引物对与探针,并制备可视芯片。通过PCR对样品核酸进行扩增,将扩增产物与固定于可视芯片的特异性探针进行杂交分析,验证了芯片的特异性和重复性,并对检测芯片灵敏度进行测试。该方法可以一次检测出5种的常见转基因植物,且实验结果表明该方法高效、准确、简便、高通量、实用性强,灵敏度可达0.1%,并摆脱了基因芯片在杂交结果分析阶段对荧光扫描仪的依赖,杂交结果明显直观。     

大豆异黄酮生理功能及其检测方法的研究进展

大豆异黄酮是大豆生长过程中形成的一类次级代谢产物,具有弱雌激素活性、抗氧化活性、抗溶血活性和抗真菌活性,能有效地预防和治疗癌症、骨质疏松、妇女更年期综合征等多种疾病。本文综述了大豆异黄酮的结构、性质及其提取、分离与分析检测方法的研究进展,拟为今后大豆异黄酮的综合利用及其衍生产品的开发提供参考。