新城疫病毒HN蛋白酵母双杂交诱饵载体的构建及鉴定

【目的】应用酵母双杂交技术构建新城疫病毒的血凝素-神经氨酸酶(HN)蛋白线性中和表位区的诱饵载体pGBKT7-HN-neu,为筛选靶向新城疫病毒HN蛋白的中和性纳米抗体奠定基础。【方法】合成HN蛋白的线性中和表位区HN-neu,将其克隆至酵母双杂交系统诱饵载体pGBKT7中,构建诱饵载体pGBKT7-HN-neu,经酶切鉴定和测序验证正确后,将pGBKT7-HN-neu转化酵母菌Y2H Gold,检测其在酵母细胞中的毒性和自激活作用。【结果】成功合成了HN-neu,构建的pGBKT7-HN-neu在酵母细胞Y2H Gold中无毒性和自激活能力。【结论】成功构建了诱饵载体pGBKT7-HN-neu。     

Acute toxicity in five dogs after ingestion of a commercial snail and slug bait containing iron EDTA

This case series of five dogs describes the effects of ingesting large amounts of an iron EDTA snail-bait product. In all cases signs of toxicity occurred between 6 and 24 h after ingestion and included abdominal pain and haemorrhagic gastroenteritis. Two of the dogs had pretreatment serum iron levels measured and in both cases the levels were above normal limits. All of the dogs were treated with iron chelation therapy and supportive care including intravenous fluids, analgesics, gastric protectants and antibiotics. Chelation therapy with desferrioxamine mesylate did not cause adverse effects in any of the dogs and all survived to discharge. The effects of iron EDTA snail bait in dogs requires further study and minimum toxic doses need to be established.     

新型软体动物毒饵研制初探

[目的]研制新型有效的软体动物毒饵。[方法]以同型巴蜗牛为试验对象,以玉米粉和豆粉为饵料,以淀粉为粘合剂,分别以白砂糖、酵母、巧克力作为引诱剂,制成诱饵,对同型巴蜗牛进行趋性试验。同时以巧克力为引诱剂,以金属络合物为有效成分,加入有玉米粉、豆粉、适量淀粉和水,制得不同有效浓度的毒饵,对同型巴蜗牛进行室内诱杀试验。[结果]对同型巴蜗牛的趋性试验表明,在3种引诱剂中,巧克力对同型巴蜗牛有较强的引诱作用 以巧克力为引诱剂制得的毒饵对蜗牛具有明显的诱杀效果,3种浓度的毒饵（3%、6%和8%）施药3 d后,同型巴蜗牛的死亡率分别达86.11%、91.67%和94.44%。[结论]用金属络合物有效成分制成的毒饵对同型巴蜗牛的毒力较高,安全性好。     

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。     

Controlling yellow jackets with fipronil-based protein baits in urban recreational areas

ABSTRACT

The western yellow jacket, Vespula penyslvanica (Saussare), is a serious seasonal pest of outdoor venues in the western United States. In the spring, queens and low numbers of workers were captured in heptyl-butyrate-baited traps until early July, when the number of foragers dramatically increased. Microsatellite data suggest that 18 colonies were actively foraging within the park in 2012. Foragers from 11 different colonies were collected at one trap site. In 2012 and 2013, sufficient numbers of foragers were not captured until early August when baiting trials were initiated. Baits were prepared with canned chicken mixed with fipronil for a final concentration of 0.025%. In 2012, a single baiting provided >96% reduction of foragers for at least two months. A second baiting late in the season provided >80% reductions. In 2013, one baiting trial resulted in a 74% and a 93% reduction. A bait acceptance study was conducted in 2014 to test a bait consisting of the juices from canned chicken and 0.025% fipronil incorporated into a hydrogel, and provided a much longer lasting bait in the field.     

用酵母双杂交筛选与南瓜蚜传黄化病毒MP互作的寄主因子

植物病毒编码的移动蛋白(movement protein,MP)介导病毒在寄主体内的移动,研究其与寄主间的分子互作有助于揭示病毒侵染过程中的分子机制。将南瓜蚜传黄化病毒Cucurbit aphid-borne yellows virus(CABYV)MP基因定向克隆到含有DNA结合功能域(DNA-binding domain,BD)载体上,并构建与激活功能域(activation domain,AD)融合表达的西瓜茎叶cDNA文库,然后用MP为诱饵筛选文库寻找与其互作的寄主因子。结果表明,诱饵质粒插入的MP基因可读框和氨基酸序列均正确,对酵母菌株AH109和Y187没有自主激活能力;文库滴度为2.94×106CFU/mL,且大多数插入片段在700bp以上,质量符合筛选要求;经过筛选和共转化回转验证,有48个候选阳性克隆与MP在酵母中互作。测序得到这些克隆的cDNA序列,BLAST分析结果表明,这些克隆共编码12种蛋白。     

Attraction of Chrysoperla carnea complex and Chrysopa spp. lacewings (Neuroptera: Chrysopidae) to aphid sex pheromone components and a synthetic blend of floral compounds in Hungary

BACKGROUND: The deployment of synthetic attractants for the manipulation of lacewing populations as aphid predators is currently used in integrated pest management. This study investigates a synthetic bait comprising floral compounds previously found to attract the Chrysoperla carnea complex, and, for the first time, the aphid sex pheromone components (1R,4aS,7S,7aR)‐nepetalactol and (4aS,7S,7aR)‐nepetalactone, in field experiments in Hungary, for their ability to manipulate lacewing populations. RESULTS: The synthetic floral bait attracted both sexes of the Chrysoperla carnea complex, and Chrysopa formosa Brauer showed minimal attraction. The aphid sex pheromone compounds alone attracted males of C. formosa and C. pallens (Rambur). When the two baits were combined, Chrysopa catches were similar to those with aphid sex pheromone baits alone, but carnea complex catches decreased significantly (by 85–88%). CONCLUSION: As the floral bait alone attracted both sexes of the carnea complex, it showed potential to manipulate the location of larval density via altering the site of oviposition. Aphid sex pheromone compounds alone attracted predatory males of Chrysopa spp. and can potentially be used to enhance biological control of aphids. For the carnea complex, however, a combination of both baits is not advantageous because of the decrease in adults attracted. Assumptions of intraguild avoidance underlying this phenomenon are discussed. Copyright © 2010 Society of Chemical Industry     

酶解啤酒废酵母生产橘小实蝇蛋白饵剂的研究

水解蛋白液是橘小实蝇蛋白饵剂的核心组成成分,研究利用酶解法从啤酒废酵母中提取水解蛋白的加工工艺。通过正交试验,对水解温度、pH、时间和加酶量4个水解影响因素进行分析,结果显示,最佳工艺条件为pH值6.0,温度50℃,时间18h,加酶量为2.0mg/mL。室内生物测定的结果表明,水解蛋白液对橘小实蝇的引诱率可达60.00%,其中雌性和雄性的引诱率分别为63.30%和56.70%。总糖与蛋白质含量的比例是影响引诱率的重要因素,当二者之比为200:1时,引诱率最高。     

复方中草药添加剂对草鱼免疫功能的影响

将大黄、黄芩、白术、党参、板蓝根等10余味中草药粉碎过筛后制成复方添加剂,按1%、2%、3%的比例添加于基础饲料中制成颗粒药饵投喂草鱼,研究复方中草药对草鱼免疫功能的影响.结果表明,草鱼血细胞吞噬率、吞噬指数和血清溶菌酶活力均显著提高;各试验组草鱼的免疫保护率也明显提高,其中以2%的添加量效果最好,能有效预防草鱼感染温和气单胞菌.     

蓝舌病病毒衣壳蛋白VP2、VP5、VP7酵母双杂交诱饵质粒的构建及鉴定

蓝舌病病毒(bluetongue virus,BTV)的结构主要由3层衣壳蛋白组成,其中VP2、VP5蛋白构成了BTV的外层衣壳,VP7蛋白构成了BTV的中间衣壳,最内层衣壳则由VP3蛋白构成。VP2、VP5及VP7蛋白在BTV侵染宿主细胞的过程中起着非常重要的作用。为了研究BTV与宿主细胞相互作用的分子机制,本研究将BTV的VP 2、VP 5、VP 7基因分别克隆到pGBKT7载体中,成功构建了pGBKT7-VP2、pGBKT7-VP5与pGBKT7-VP73个诱饵质粒,且通过自激活和毒性验证,证明所构建的3个质粒均无自激活作用,对酵母细胞无毒性作用。本研究为今后利用酵母双杂交筛选VP2、VP5、VP7蛋白中与宿主细胞相互作用的蛋白做好了铺垫,为深入研究BTV与宿主细胞的相互作用奠定了基础。