猪轮状病毒Vp4全基因与乳酸菌非抗性表达载体的重组及在大肠杆菌中的表达

以猪A组轮状病毒mRNA为模板，应用优化的反转录聚合酶链式反应（RT—PCR）技术，扩增了2331 bp的Vp4全长基因，通过T-A克隆技术，将PCR产物克隆至克隆载体pGEM-T中，构建质粒pGEM-T-Vp4。以SalⅠ和KpnⅠ双酶切pGEM—T—Vp4．和以胸苷酸合成酶基因（thymidylate synthase，thyA）为选择压力的非抗生素抗性的穿梭表达载体pW425t，并将纯化的Vp4基因亚克隆至表达载体pW425t中，构建出可以在乳酸菌与大肠杆菌之间穿梭表达的原核表达重组质粒pW425t-Vp4。将pW425t-Vp4转化至thyA基因缺陷型的大肠杆菌感受态E．coli X13中，通过生长功能弥补筛选阳性克隆，SDS-PAGE分析，可见约87.67Ku融合蛋白。Western blot分析表明，该蛋白具有与轮状病毒多克隆抗体的反应原性，研究结果为pW425t-Vp4在乳酸菌受体菌株中表达提供科学依据。     

城市空气质量评价中的模糊支持向量机方法

传统的评价方法不能处理带有模糊信息的城市空气质量的评价问题。基于结构风险最小化原则建立了带有模糊信息的支持向量分类模型和算法,在此基础上提出基于模糊支持向量机的城市空气质量评价方法;根据空气中的总悬浮颗粒物、二氧化硫、二氧化氮、空气综合污染指数4个指标,对我国50个主要城市2003年空气质量进行了综合评价,选择10个城市的评价结果进行了测试,结果表明:根据2003年数据采用本方法得到的邢台、张家口和沧州的空气污染指数与河北省环保专家提供的数据误差小于0.1;西宁、银川、乌鲁木齐、汕头、南京、大连、武汉的综合空气污染指数与国家环保总局提供的数据误差均小于0.5。     

基于MK—SVR模型的小麦叶面积指数遥感反演

提出了运用多核支持向量回归(MK-SVR)算法构建小麦叶面积指数(LAI)遥感监测模型。以2010—2013年试验样点小麦拔节、孕穗、开花3期的实测LAI数据为基础,同步获取我国自主研发的环境减灾卫星HJ-CCD对该研究区域的影像数据,分析了各生育期小麦LAI与8种植被指数间的相关性。以显著相关的植被指数作为输入参数,使用MK-SVR算法构建了每个生育期的小麦LAI反演模型,即MK-SVR-LAI模型。为了评价模型,每期使用单一核支持向量回归(SK-SVR)、偏最小二乘(PLS)回归算法构建了SK-SVR-LAI、PLS-LAI模型。将模型估算LAI值和田间观测LAI值进行比对,以决定系数(R2)和均方根误差(RMSE)为指标评价并比较了模型。结果表明:3个生育期MK-SVR-LAI模型的RMSE值均低于参比模型,拔节期为0.293 1,孕穗期为0.466 8,开花期为0.548 6,且该模型的R2也都最高,拔节期为0.762 4,孕穗期为0.801 8,开花期为0.668 9。     

利用透明颤菌血红蛋白基因vgb改造毕赤酵母分泌型表达载体pPIC9K的研究

为了解决酵母发酵时贫氧限制目的蛋白表达量,利用透明颤菌血红蛋白基因vgb设计毕赤酵母分泌 型表达载体pPIC9K-vgb,并以人乳铁蛋白基因为目的基因转入毕赤酵母GS115。研究结果表明,该表达载体具 有转化操作简单、后期工程菌筛选简便、菌体耐低氧能力强、目的蛋白摇瓶表达量高等优点。该表达载体也可 以通过直接替换目的基因而用于其他蛋白表达系统的开发研究。     

Disease progression of a lethal decline caused by the 16SrIV‐D phytoplasma in Florida palms

Recently, a new phytoplasma was discovered in Hillsborough County in the state of Florida, USA. This phytoplasma belongs to the 16SrIV taxonomic group and is classified as subgroup D. It is the causal agent of lethal bronzing disease (LBD) of palm. Since the discovery of LBD in 2006, the disease has spread throughout much of the state. In 2014 and 2015, stands of cabbage palm and queen palms that had been present at the University of Florida's Fort Lauderdale Research and Education Center in Davie, FL began showing symptoms of LBD. After confirming the presence of the LBD phytoplasma in initially infected palms by nested PCR and RFLP analysis, all palms were systematically sampled over the period of 1 year to monitor and quantify disease spread. A total of 30 cabbage palms were tested monthly by qPCR, with five testing positive on the first sample date. By the end of the study period, 16 cabbage palms had died from the infection. A total of 16 queen palms were surveyed, with three palms initially testing positive. By the end of the study, four queen palms had tested positive and died from the infection. To the authors’ knowledge, this study is the first to document and quantify spread of palm‐infecting phytoplasmas. This data provides important insights into the ecology of palm‐infecting phytoplasmas and highlights the impact that the movement of infective insects can pose to established stands of palms.     

流产型布鲁氏菌Omp22基因酵母双杂交诱饵载体的构建及鉴定

通过PCR获得流产型布鲁氏菌Omp22基因的编码区,将其克隆到pSOS载体中,构建酵母双杂交系统的诱饵载体pSOS-Omp22。测序正确后,将重组质粒导入cdc25H检测其表达产物对酵母细胞有无毒性和自激活作用,且在酵母细胞中定位是否正确。结果表明,获得了正确的流产型布鲁氏菌Omp22基因编码区,并成功克隆到pSOS诱饵载体中,且转化有诱饵载体的cdc25H在SD/Glucose(-L)营养缺陷平板上生长良好,说明表达产物对酵母细胞无毒性,对报告基因也无自激活作用且其定位正确。表明pSOS-Omp22可应用在酵母双杂交系统中,用于寻找巨噬细胞cDNA文库中与Omp22相互作用的蛋白质。     

拟南芥AtHOS10基因的克隆及其表达载体的构建

AtHOS10基因对植物冷调节起到重要作用,可以通过控制ABA合成来改变植物的脱水胁迫耐性。本文利用PCR方法从Columbia生态型的拟南芥基因组中扩增出冷调节基因AtHOS10,并插入克隆载体。序列分析表明,克隆片段长3453bp,与目前发表的序列完全一致。将pUC-AtHOS10和pBI-HEM进行酶切重组构建表达载体pBI-AtHOS10,然后将表达载体转化导入农杆菌中,为侵染烟草作准备。     

基于局部二值模式和灰度共生矩阵的籽棉杂质分类识别

籽棉杂质的分类识别是实现棉花生产线自适应加工的基础与重要依据。该文提出了一种基于局部二值模式和灰度共生矩阵的籽棉杂质分类识别算法,该算法将含杂籽棉图像首先转换为局部二值模式图像,获取图像的微观结构,再用局部二值模式图像生成灰度共生矩阵并计算特征参数,获取图像宏观结构。使用支持向量机作为分类器,用不同尺度的图像结构进行训练,从而达到籽棉杂质的分类识别。试验结果表明,该文设计算法对各种杂质的平均正确识别率达到了94%,超过单独使用局部二值模式和单独使用灰度共生矩阵的正确识别率,为实现棉花自适应加工提供了技术基础。     

Experimental comparison of aerial larvicides and habitat modification for controlling disease-carrying Aedes vigilax mosquitoes

BACKGROUND: Microbial and insect‐growth‐regulator larvicides dominate current vector control programmes because they reduce larval abundance and are relatively environmentally benign. However, their short persistence makes them expensive, and environmental manipulation of larval habitat might be an alternative control measure. Aedes vigilax is a major vector species in northern Australia. A field experiment was implemented in Darwin, Australia, to test the hypotheses that (1) aerial microbial larvicide application effectively decreases Ae. vigilax larval presence, and therefore adult emergence, and (2) environmental manipulation is an effective alternative control measure. Generalised linear and mixed‐effects modelling and information‐theoretic comparisons were used to test these hypotheses. RESULTS: It is shown that the current aerial larvicide application campaign is effective at suppressing the emergence of Ae. vigilax, whereas vegetation removal is not as effective in this context. In addition, the results indicate that current larval sampling procedures are inadequate for quantifying larval abundance or adult emergence. CONCLUSIONS: This field‐based comparison has shown that the existing larviciding campaign is more effective than a simple environmental management strategy for mosquito control. It has also identified an important knowledge gap in the use of larval sampling to evaluate the effectiveness of vector control strategies. Copyright © 2011 Society of Chemical Industry     

家蚕抗菌肽基因Cecropin-D的克隆及其真核表达载体的构建

从家蚕中提取总RNA,采用反转录聚合酶链式反应(RT-PCR)方法,获得了家蚕Cecropin-D基因编码区的cDNA,扩增出家蚕抗茵肽Cecropin-D成熟肽基因片段,重组克隆入pMD18-Tvector载体,经DNA测序,该基因为186bp,编码62个氨基酸。用限制性内切酶切下目的基因,插入毕赤酵母表达载体pPIC9K中,构建真核表达栽体pPlC9K-CD。本实验的研究为获得超量表达的高活性表达抗菌肽Cecropin-D以及为研制具有抗菌活性的新型基因药物奠定了基础。