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以伪狂犬病病毒GB株细胞培养物为模板，用PCR方法扩增蛋白激酶(PK)基因，对其进行了克隆、序列测定，并与PRV NIA-3株、Ka株和其他α-疱疹病毒进行了同源性比较。结果显示：所扩增的PK基因片段长1396bp，该片段包含一个开放阅读框(ORF)，长1005bp，编码由334个氨基酸组成的蛋白质。与NIA-3株及Ka株的核苷酸同源性为98．4％和97．5％，基因编码区氨基酸序列的同源性为98．2%和96．4％。具有真核细胞丝氨酸／苏氨酸蛋白激酶氨基酸亚结构域特征序列，而且α-疱疹病毒PK基因具有一些特征性的、共有的氨基酸序列。为今后研究PK基因的功能和构建PK缺失株奠定了基础。 相似文献
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本文对33株枯草芽孢杆菌群菌株进行β-甘露聚糖酶活性筛选,其中的32株具有β-甘露聚糖酶活性,只有1株无β-甘露聚糖酶活性。通过基因克隆测序的方法获得33株枯草芽孢杆菌群菌株β-甘露聚糖酶基因编码区全序列,对酶基因进行同源性分析并构建系统发育树;在β-甘露聚糖酶基因系统发育树中,33株枯草芽孢杆菌群菌株聚为3个分支,分别是枯草芽孢杆菌分支、地衣芽孢杆菌分支和解淀粉芽孢杆菌分支;枯草芽孢杆菌、地衣芽孢杆菌和解淀粉芽孢杆菌β-甘露聚糖酶基因种内同源性大于91％,而种间同源性为60％～69％。 相似文献
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嗜水气单胞菌aer毒素基因的克隆及核苷酸序列分析 总被引：2，自引：0，他引：2
以自行分离的嗜水单胞菌分离株AHCS02为研究对象,分析其产生的aer毒素基因结构.根据GenBank中aer毒素的基因序列（M16495）,设计扩增编码aer毒素成熟蛋白基因的引物,以AHCS02的基因组DNA为模板,PCR扩增出长度为1 332 bp的核苷酸片段,进行pMD18-T载体克隆.酶切鉴定后进行序列测定和分析,结果表明扩增的片段与M16495的同源性达92%,将测得的序列登录GenBank数据库,所得的序列编号为AY136943.分析推导其核苷酸编码的氨基酸序列显示：其编码443个氨基酸,为aer毒素成熟蛋白的基因,与M16495相比,氨基酸差异主要集中在422氨基酸残基至436氨基酸残基的位置上,在这个位置上共有8个氨基酸残基发生变化,其中在435和437氨基酸残基之间有一个氨基酸密码子丢失,氨基酸的同源性达95.5%.证明aer毒素基因的5＇端保守,3＇端变异较大. 相似文献
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环介导等温扩增技术对牛奶中沙门氏菌快速检测的研究 总被引：1，自引：0，他引：1
环介导等温扩增技术(LAMP)是一种新型的基因扩增方法,它具有高特异性、高灵敏度、便捷、成本低等特点,因而广泛应用于临床检测.本实验以沙门氏菌高度保守基因fimY基因为靶基因设计两对引物(F3&B3、FIP&BIP),建立LAMP反应体系.经试验分析对体系特异性、Mg2+浓度、反应时间、反应温度进行了优化和摸索,对35株临床菌株均检出达100％,其他阴性菌株基因组均无检出,在人工添加样品检测得其在牛奶中灵敏度为33CFU/管.实验结果表明,该方法耗时少、具有高特异性,适合牛奶中沙门氏菌污染的快速检测. 相似文献
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从土样中筛选获得一株α－淀粉酶产量的芽孢杆菌菌株,编号为ＸＤ９１３。通过克隆和测序,得到了一个２０９０ｂｐ的ＤＮＡ片段,内含一个编码５１４个氨基酸的基因,经同源性比较,发现该基因与已发表的解淀粉芽孢杆菌的α－淀粉酶基因的同源性高达９９．８％。 相似文献
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采用以结晶纤维素（Avicel）为唯一碳源的平板活性筛选法,从60份森林腐殖土、腐术样品中分离得到2株在pH5．0、37℃条件下高效降解结晶纤维素的细菌菌株GXN152和GXN153。以菌株的总DNA基因为模板,用细菌的16S rRNA基因的通用引物扩增得到菌株GXN152和GXN153的16S rRNA基因序列。测序分析表明菌株GXN152的16S rRNA基因序列和洋葱假单胞菌（Burkholderia cepacia）菌株CNR22的16S rRNA基因序列的同源性最高,具有98％的同源性,生理生化特性检测表明菌株GXN152具有洋葱假单胞菌的鉴别特征,将结晶纤维素降解菌GXN152鉴定为洋葱假单胞菌。菌株GXN153的16S rRNA基因序列和类芽孢杆菌（Paenibacillus favisporus）菌株GMP01的16S rRNA基因序列的同源性最高,具有99％的同源性,生理生化特性检测表明菌株GXN153具有类芽孢杆菌的鉴别特征,将纤维素降解菌GXN153鉴定为类芽孢杆菌。 相似文献
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用PCR扩增猪圆环病毒II型广东分离株的衣壳蛋白羧基端基因,将PCR产物连接到重组型质粒pR上,转化DH5α细胞并筛选阳性克隆;重组质粒经PCR鉴定并测序后,转化E2菌,将重组E2菌与缺陷型噬菌体T4-Z1同源重组后,得到重组噬菌体,SDS-PAGE和Westernbloting分析,表明衣壳蛋白基因在噬菌体表面正确展示,表达的融合蛋白的分子量约为25Ku。 相似文献
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为调查广西猪戊型肝炎（HE）的感染情况,本研究应用套式RT-PCR方法对采自广西14市43个猪场的508份3月龄左右猪只粪样及市售182份猪肝扩增猪戊型肝炎病毒（HEV）ORF2基因特异序列,并进行核苷酸序列同源性分析及遗传进化分析。结果表明,在508份猪粪样中194份可以检测到HEV RNA,阳性率为38.2%;182份猪肝样本中,33份可检测到HEV RNA,阳性率为18.1%;43个猪场中检出HEV RNA的有35个,猪场阳性率为81.4%。所获取的39株广西猪HEV ORF2基因特异序列同源性为89.9%～100%,提示在所调查的广西猪场中已较普遍存在HEV感染,所获取的39株广西HEV阳性样品均属于基因Ⅳ型。本研究结果佐证了感染猪及市售带毒猪肝脏是HEV重要的储存库和传染源,应该重视其公共卫生学意义,预防粪-口途径或者食源性HEV感染。 相似文献
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根据GenBank中猪圆环病毒2型(Porcine circovirus2,PCV2)的核苷酸序列设计2对引物,使用扩增全基因的引物对来源于河南省焦作、开封和滑县的3株PCV 2型JZ株、KF株和HX株的ORF2基因进行扩增,扩增片段克隆到pMD18 T上,获得重组质粒pMD18-T-ORF2,并对其进行测序,测序结果登录在GenBank上,登录号分别为EF028202、EF064149和EF467928.序列分析表明,PCV2河南分离株ORF2基因与其它PCV2 ORF2基因核苷酸序列同源性为92.4%～99.6%,氨基酸序列同源性为90.6%～97.9%,进化分析表明,河南分离株处于同一分支,与欧洲株亲缘关系较近.应用另一对引物从重组质粒pMD18-T-ORF2中PCR扩增出587 bp不包含核定位信号序列的ORF2基因,克隆到表达载体pET-32a,成功构建了重组质粒pET-32a-ORF2,经IPTG诱导表达了ORF2基因编码的结构蛋白,经SDS-PAGE和Western blotting检测,表达的重组蛋白分子量约为40 kD,可被PCV2阳性血清识别,说明PCV2 ORF2基因得到成功表达. 相似文献
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Physicochemical properties of β and α'α subunits isolated from soybean β-conglycinin 总被引：1，自引：0，他引：1
Soy protein has shown great potential for use in biobased adhesives. β-Conglycinin is a major component of soy protein; it accounts for 30% of the total storage protein in soybean seeds. β-Conglycinin was isolated and purified, and its subunits' (β, α'α) physicochemical and adhesive properties were characterized. Crude β-conglycinin was isolated from soy flour and then purified by the ammonium sulfate precipitation method. The α'α and β subunits were isolated from the purified β-conglycinin by anion exchange chromatography. Yields of α'α subunits and β subunits from 140 g of soy flour were 1.86 g (1.3%) and 0.95 g (0.67%), respectively. The minimum solubility for α'α subunits, β subunits, and β-conglycinin occurred in pH ranges of 4.1-5.4, 3.5-7.0, and 4.8-5.3, respectively. Transmission electron microscopy showed that the β subunits existed as spherical hydrophobic clusters, whereas α'α subunits existed as uniformly discrete particles at pH 5.0. Differential scanning calorimetry showed that β subunits had higher thermal stability than α'α subunits. The pH had a lesser effect on adhesion strength of the β subunits than on that of the α'α subunits. The adhesives made from β subunits also showed greater water resistance than those from α'α subunits and β-conglycinin. Soy protein rich in β subunits is likely a good candidate for developing water-resistant adhesives. 相似文献
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淡水鱼腹背定向装置设计及试验 总被引：4，自引：4，他引：0
针对目前淡水鱼前处理设备主要采用人工进行定向喂料,劳动强度大、效率低,同时鱼体定向整理设备缺乏等问题,该研究以鱼体腹部横截面近似为楔形的典型大宗淡水鱼鲫鱼为研究对象,在鱼体腹背受力挤压力学特性研究的基础上,提出了一种淡水鱼鱼体腹背定向方法,同时分析了鱼体腹背定向机理,设计并试制了淡水鱼鱼体腹背定向试验装置。选取鲫鱼为试验对象,以定向成功率和定向时间为试验指标,对淡水鱼鱼体腹背定向试验装置的主要参数进行了优化。通过单因素试验探究了辊形、对辊间隙、鱼体输送速度、对辊转速等装置主要参数对鱼体腹背定向的影响规律。在单因素试验结果的基础上,进行了正交试验,试验结果表明:各因素间的最优水平组合为:对辊间隙50 mm,对辊转速50 r/min,鱼体输送速度0.6 m/s时,在此最优水平组合下,定向成功率为100%,定向时间为0.55 s,试验装置工作运行参数最佳。该文的研究结果可为淡水鱼鱼体定向设备的研发提供理论依据和技术参考。 相似文献
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Peter S. Maitland 《Biological conservation》1974,6(1):7-14
This paper briefly reviews the status and distribution of populations of freshwater fishes in the British Isles and their worth as a resource. The total value of freshwater fishes to the community is far greater than is normally appreciated and includes, in addition to the economic sport and commercial fisheries, amenity, recreational, educational, and scientific, aspects as well as a potentially useful store of genetic material for the future. The main human pressures on existing fish stocks are discussed under the headings of fisheries, pollution, and land-used. The principal trends in the British Isles are away from natural and stable mixed fish populations towards artificially maintained, unstable stocks of a few species of sporting or commercial value. In particular the rarer, more sensitive, fish stocks with poor powers of distribution are being eradicated and replaced by commoner, more robust forms with greater powers of distribution. The rarer species and genetic strains can only survive if constructive conservation projects are initiated expeditiously at a variety of levels. 相似文献
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Human development of freshwater ecosystems has lead to drastic changes in freshwater fish faunas, including the loss of many native species and the gain of non-natives. Typically conservation ecologists view these two opposing forces as contributing to biological homogenization, and consider homogenization as one of the principle negative consequences of urbanization. However, homogenization is only one outcome out of many that can result from the loss and gain of species. In particular, it is possible for invasions and extinctions to lead to differentiation; a process whereby two (or more) regions become less similar to one another through time. Using the freshwater fishes of California, we show that urbanization is highly positively correlated to both the endangerment of native fish and the invasion of non-native fish within watersheds. Despite this, the fish faunas of California’s watersheds have differentiated from one another through time. Furthermore, the degree of differentiation is positively correlated with measures of urbanization, which is contrary to expectation. We suggest that this result reflects: (1) the haphazard manner in which non-native fishes have been introduced into California watersheds, (2) the difficulty that both native and non-native fishes have in expanding their geographical ranges, and (3) the continued presence of vestiges of formerly distinct regional faunas. This pattern of differentiation among watersheds is likely a matter of scale, as previous work on freshwater fishes has demonstrated homogenization at both larger and smaller spatial scales. In addition the observed pattern is probably a short-term (temporal) phenomena and will disappear with continued invasion and extinction. We suggest that similar patterns may occur for other taxa that have limited natural dispersal abilities and that are idiosyncratically released as non-natives via human activities (e.g. herptiles). 相似文献
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Moreno IM Maraver J Aguete EC Leao M Gago-Martínez A Cameán AM 《Journal of agricultural and food chemistry》2004,52(19):5933-5938
The presence of cyanobacterial toxins (microcystins) in waters and food increases the risk of toxicity to animal and human health. These toxins can degrade in the human gastrointestinal tract before they are absorbed. To evaluate this possible degradation, water samples spiked with known concentrations of microcystins MC-LR, MC-RR, and MC-YR, which are the toxins most commonly produced by such toxic cyanobacteria as Microcystis aeruginosa, Oscillatoria spp., and Nostoc spp., were submitted to a dissolution test that used gastric and intestinal fluids according to U.S. Pharmacopeia conditions. HPLC with UV detection was used to determine the toxins before and after treatments. This study revealed enzymatic alterations in gastric conditions for all the toxins assayed. MC-RR was the toxin most affected: its range of inactivation was 49-64%. The percentage of degradation for MC-YR and MC-LR was around 30%. However, none was degraded by intestinal digestion. 相似文献
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通过分析广州市区居民消费量较大的7种淡水鱼和5种海水鱼肌肉中甲基汞（MeHg）含量,评估人体通过食鱼摄入MeHg的暴露风险。结果显示,广州市鱼体肌肉中MeHg含量范围为2.39～349μg·kg-1,平均含量为43.4μg·kg-1。不同食性鱼类MeHg含量水平依次为：肉食性〉杂食性〉草食性,处于食物链不同环节解释了这种含量差异。从生长环境来看,海水鱼肌肉的MeHg含量总体上高于淡水鱼。所有鱼中MeHg含量均低于国内外相关标准限值。与国内其他地区商品鱼相比,广州市商品鱼中MeHg含量相当,但明显低于野生鱼。居民通过食鱼摄入的MeHg范围为0.035～0.087μg·kg-1·d-（1bw）,均值为0.050μg·kg-1·d-（1bw）,远低于美国EPA规定的MeHg暴露参考剂量（RfD）值（1.1μg·kg-·1d-1）,也低于WHO和FAO制定的临时性周可承受摄入量（PTWI）（1.6μg·kg-1）。研究结果表明,广州居民食鱼MeHg暴露风险较小,但儿童等敏感人群食用特定鱼类的量和频次要引起一定注意。 相似文献
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D A Kautter H M Solomon 《Journal of the Association of Official Analytical Chemists》1977,60(3):541-545
The mouse toxicity and protection technique for the detection and identification of Clostridium botulinum and its toxins in foods was collaboratively studied by 11 laboratories. Each laboratory received 4 samples of cream of mushroom soup; 2 contained spores and toxin of C. botulinum type A, 1 contained spores and toxin of C. botulinum type E, and 1 contained spores of C. sporogenes. The media used were cooked meat medium (beef heart or chopped liver broth) and trypticase peptone glucose yeast extract broth with trypsin. The results indicate that this method has a high degree of repeatability and reproducibility. All 11 laboratories correctly identified the toxins and the nontoxic sample in the food and detected and identified the viable spores in the samples by means of the subsequent cultures. This method has been adopted as official first action. 相似文献
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The stability, crystallization, and melting behavior of canola stearin (CaSt) solid lipid nanoparticle dispersions (SLN) and canola oil-in-water emulsions (COE) with 10 wt % Poloxamer 188 (P188) or Tween 20 (T20) with and without 0.1 wt % β-carotene (BC) were investigated. Particles or droplets with diameters in the range of 115 nm were formed and stable for up to 90 days at 4 or 20 °C. Polymorphism was affected by surfactant type; that is, only β versus both β' and β were observed for the P188 and T20 SLN, respectively. According to Cryo-TEM, the emulsions and SLN were spherical versus platelet-like structures, respectively, with differences observed between SLN with P188 or T20. More surfactant was interfacially adsorbed in the SLN versus COE. Incorporation of BC at 0.1 wt % had no impact on SLN or COE size, polymorphism, or melting behavior. Less BC degradation was observed for the SLN versus COE and during storage at 4 versus 20 °C (p < 0.05). 相似文献
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Boselli E Pacetti D Lucci P Frega NG 《Journal of agricultural and food chemistry》2012,60(12):3234-3245
The phospholipid molecular species of freshwater (pangasius, Nile perch, trout), marine fish fillets (horse mackerel, European hake, common sole, European anchovy, European pilchard, Atlantic mackerel) and the edible muscle foot of bivalves (clam, mussel, oyster) commonly available in the Italian market during spring and summer were characterized by means of normal-phase high performance liquid chromatography coupled online with positive electrospray ionization ion-trap tandem mass spectrometry. From principal component analysis (PCA), it was observed that the total fatty acid profile was not suitable to differentiate among the shellfish genera. The fatty acid molecular combinations of phosphatidylcholine, the main phospholipid class, as well as phosphatidylinositol and phosphatidylethanolamine allowed for the differentiation of shellfish from the bony fishes. Phosphatidylserine and phosphatidylethanolamine plasmalogen profile allowed for the discrimination of each bony fish or shellfish genus since PS and pPE classes included a large number of fatty acid combinations that were specific for a fish genus or group. 相似文献