2个抗虫棉的外源Bt基因分子鉴定及其染色体定位

以中美2个抗虫棉品种GK19与33B为试验材料,利用检测中美Bt基因的特异性引物,分别对抗虫棉亲本GK19和33B进行PCR扩增,并通过SSR分子标记技术对其Bt基因进行分子鉴定与染色体定位,旨在从外源基因转化事件的视角探究中美转基因抗虫棉差异的分子基础。结果表明, GK19为中国转Bt基因抗虫棉, 33B为美国转Bt基因抗虫棉; GK19的Bt基因被定位在棉花Chr.20上,共16对SSR多态性标记与其Bt基因连锁,两侧的分子标记为NAU3907和NAU2579,其遗传距离分别为2.4 cM和1.5 cM; 33B的Bt基因被定位在棉花Chr.26上,共20对SSR多态性标记与Bt基因连锁,目标Bt基因位于标记NAU460和dc40260之间,其遗传距离分别为3.6 cM和2.0 cM。以上结果表明GK19和33B属于不同的遗传转化事件。     

木尔坦棉花曲叶病毒编码蛋白的亚细胞定位分析

 木尔坦棉花曲叶病毒(Cotton leaf curl Multan virus,CLCuMV)是典型的单组分双生病毒,并伴随β卫星分子,是棉花曲叶病的主要病原之一。本研究利用农杆菌介导的瞬时表达系统,将CLCuMV及其卫星分子编码的7个病毒蛋白在本氏烟表皮细胞中表达。通过激光共聚焦显微镜观察发现:V1、C2和C3定位于细胞核;C1和βC1定位在细胞核以及细胞质或细胞膜上,并且在细胞质中形成丝状结构;V2和C4主要定位在细胞质或细胞膜上,细胞核也有微量表达,V2可形成大小不一的颗粒状聚集体结构,C4在细胞膜上可见点状聚集体结构。同时,利用RT-PCR和Western blot对病毒各基因的转录和表达水平进行了分析。CLCuMV编码蛋白的亚细胞定位为蛋白功能的进一步研究提供重要理论依据。     

RTK测量系统在古树名木保护中的应用

RTK测量技术是GPS测量技术与数据传输技术的结合,可实时获得高精度三维定位信息,是测量领域又一次技术革命。本文以四方井水库古树名木保护定位测量为例,阐述RTK的含义、系统组成,分析RTK测量系统的优点以及在测量过程中应注意的问题,为提高林业勘测效率和结果准确性提供参考。     

Accuracy of the TurfTrax Racing Data System for determination of equine speed and position

Reasons for performing study: The speed and position data collected by TurfTrax Racing Data Limited during UK Thoroughbred racing have potential to benefit equine science and welfare. The size (the 2006 data set alone consists of 30,932 individual horse starts across 2667 races) and nature (speed and 2D position for each horse at 4 updates per second) of the data make it a unique resource for questions in equine safety, welfare, performance, and animal locomotion. Objective: To determine the accuracy of the TurfTrax tracking system in estimating the speed and position of horses during racing. Methods: Measurements from the TurfTrax wireless tracking system were compared with those of a survey‐grade global positioning system (GPS) receiver. Results: The TurfTrax system was found to give position measurements to within ± 11 and ± 64 cm in the fore‐aft and lateral directions, respectively, averaging ± 38 cm (interquartile range) and speed to within 0.15 m/s. Potential relevance: The data collected by the TurfTrax system are of sufficient accuracy to inform new diagnoses, training regimens and basic locomotor scientific studies. The position data can provide the precise distance, going, inclination, rate of turn and pack positioning through which each horse has raced. The speed profile can be used to examine the level of exertion, effect of training regimens and influence of racecourse features on performance. A first clinical application would be to analyse retrospectively these factors on occurrence of injury to compare with past training regimens, levels of exertion, and/or racecourse conditions.     

日本结缕草ZjNAC1的克隆、转录激活活性、亚细胞定位及表达分析

采用RACE技术从日本结缕草中克隆得到ZjNAC1转录因子(GenBank No. MH428376),其开放阅读框为945bp,编码314个氨基酸。ZjNAC1编码的蛋白在N端有1个典型的NAM保守结构域,属于NAC转录因子家族。通过染色体步移的方法,获得ZjNAC1基因ATG上游1635bp序列,分析发现其包含响应脱落酸(ABA)、茉莉酸甲酯(MeJA)及逆境胁迫的作用元件。构建pGBKT7-ZjNAC1载体,转化Y2HGold酵母感受态细胞,发现ZjNAC1具有转录激活活性。农杆菌介导的35S-ZjNAC1-YFP载体注射本生烟草叶片瞬时表达结果显示,ZjNAC1定位于细胞核。实时荧光定量结果表明,ZjNAC1在日本结缕草根中表达量最高;此外,ZjNAC1的表达受10μmol/L MeJA和20%PEG4000处理的诱导,但受200μmol/L乙烯(ET)、10μmol/L ABA和300mmol/L NaCl处理的抑制。     

Identification of a cell-wall peptidase (NlpC/P60) from Nocardia seriolae which induces apoptosis in fathead minnow cells

Nocardia seriolae, a Gram-positive bacterium, is the main pathogen of fish nocardiosis. Protein NlpC/P60 is a cell-wall peptidase and a potential virulence factor of N. seriolae. Subcellular localization research revealed that both NlpC/P60-GFP and NlpC/P60Δsig-GFP fusion proteins were evenly distributed in the whole cell of fathead minnow (FHM) cells. Furthermore, typical apoptotic features, such as nuclear pyrosis and apoptotic bodies, were observed in the transfected FHM cells and grouper spleen cells by the overexpression of protein NlpC/P60. Then, quantitative assays of mitochondrial membrane potential (ΔΨm) value, caspase-3 activity and apoptosis-related gene (Bax, BNIP3, TNF1 and TNF6) mRNA expression were conducted. The results showed that ΔΨm was decreased, caspase-3 was significantly activated, and the mRNA expression of pro-apoptotic genes (Bax and BNIP3) and tumour necrosis factors (TNF1 and TNF6) was up-regulated in NlpC/P60-overexpressed cells. Taken together, the results indicated that the protein NlpC/P60 of N. seriolae might involve in apoptosis regulation. This study may lay the foundation for further study on the function of N. seriolae NlpC/P60 and promote the understanding of the virulence factors and pathogenic mechanism of N. seriolae.     

Identification of a mitochondrial‐targeting secretory protein from Nocardia seriolae which induces apoptosis in fathead minnow cells

Nocardia seriolae is the main pathogen responsible for fish nocardiosis. A mitochondrial‐targeting secretory protein (MTSP) 3141 with an N‐terminal transit peptide (TP) from N. seriolae was predicted by bioinformatic analysis based on the genomic sequence of the N. seriolae strain ZJ0503. However, the function of the MTSP3141 and its homologs remains totally unknown. In this study, mass spectrometry analysis of the extracellular products from N. seriolae proved that MTSP3141 was a secretory protein, subcellular localization research showed the MTSP3141‐GFP fusion protein co‐localized with mitochondria in fathead minnow (FHM) cells, the TP played an important role in mitochondria targeting, and only the TP located at N‐terminus but not C‐terminus can lead to mitochondria directing. Moreover, quantitative assays of mitochondrial membrane potential (ΔΨm) value, caspase‐3 activity and apoptosis‐related gene (Bcl‐2, Bax, Bad, Bid and p53) mRNA expression suggested that cell apoptosis was induced in FHM cells by the overexpression of both MTSP3141 and MTSP3141ΔTP (with the N‐terminal TP deleted) proteins. Taken together, the results of this study indicated that the MTSP3141 of N. seriolae was a secretory protein, might target mitochondria, induce apoptosis in host cells and function as a virulence factor.     

鸡Toll样受体15单克隆抗体的制备及其初步应用

旨在制备鸡Toll样受体15 (ChTLR15)的特异性单克隆抗体(mAb),并初步应用.利用PCR技术扩增ChTLR15第162-386位氨基酸,克隆于载体pET-30a中进行诱导表达,获得高纯度的重组ChTLR15 (162-386 aa)蛋白.然后采用皮下多点注射方式将ChTLR15免疫6周龄的雌性BALB/c小...     

基于探地雷达杂波抑制与偏移成像的树木根系定位方法

探地雷达在果树和古树名木养护管理领域具有广阔的应用前景。针对探地雷达采集的B-scan图像中杂波影响根系定位精度的问题,提出了基于鲁棒深度自动编码器(RDAE)、直接最小二乘法(DLS)和频率-波数域偏移(FKM)相结合的树木根系定位方法。首先,通过RDAE将零点校正后的B-scan图像分解为表示杂波的低秩分量和表示根系目标回波的稀疏分量,保留稀疏分量完成杂波抑制;然后使用DLS拟合目标回波形成的双曲线估算土壤的相对介电常数;最后,根据土壤的相对介电常数计算得出偏移速度作为频率-波数域偏移的输入进行偏移成像,获取根系的半径和深度信息从而完成根系定位。实验结果表明：RDAE方法在仿真和实测数据上的杂波抑制效果对比均值减法(MS)、奇异值分解(SVD)和鲁棒主成分分析(RPCA)有着更高的信杂比和改善因子,通过DLS估计的土壤相对介电常数均方根相对误差为3.84%,根系定位的最大半径相对误差和最大深度相对误差分别为8.5%、8.7%,能够完成根系位置标定,满足根系无损检测的需求,可为树木健康管理和移植提供决策支持。