环境水体中GⅡ型诺如病毒浓缩研究

自行设计一种适合野外现场采样的病毒采集浓缩仪,采用一种新型阳离子膜NanoCeram,进行环境水体中GⅡ型诺如病毒浓缩研究。通过预实验,考察了预处理膜、不同的二次浓缩方法、不同洗脱液对病毒回收率的影响,随后优化整个浓缩过程并确定最佳的病毒浓缩方法。结果显示,预处理膜对病毒回收影响较大,PEG-NaCl沉淀、硅藻土吸附这两种二次浓缩方法的效果相当,选择0.15mol/L Na2HPO4作为硅藻土洗脱液。确定了NanoCeram阳离子膜的病毒回收率为3.02%。最后对北京市丰台区洋桥生活污水进行现场采样,成功检测到GⅡ型诺如病毒,证明该方法有效可行,适合于环境水体中诺如病毒的浓缩分离和检测。     

胸腺肽α1-Spl DnaX内含肽融合蛋白的切割动力学研究

目的:研究胸腺肽α1(Tα1)与Spl DnaX内含肽融合蛋白AS的体外切割动力学。方法:构建Tα1和SplDnaX内含肽的融合表达载体pET-AS,转化大肠杆菌BL21(DE3),经乳糖诱导获得可溶性表达的融合蛋白AS,用镍亲和层析纯化该蛋白;综合评价温度、β-巯基乙醇(β-ME)浓度和诱导切割时间对Spl DnaX内含肽介导融合蛋白AS自切割释放Tα1的影响。结果:在大肠杆菌中诱导表达了融合蛋白AS,经镍柱纯化获得该蛋白;随着温度升高和β-ME浓度增加,诱导切割时间延长,Spl DnaX内含肽介导的切割率逐渐增大;最终采用300 mmol/Lβ-ME切割24 h,融合蛋白的硫解切割率大于90%。结论:通过对Spl DnaX内含肽的诱导切割条件进行摸索,确定了最适宜的切割条件,为利用该方法制备Tα1和其他小分子多肽提供了技术基础。     

准噶尔小胸鳖甲抗冻蛋白MpAFP698在真核细胞中的表达与鉴定

目的:构建真核表达质粒pEGFP-N1-mpafp698,转染人胚肾293T细胞,并表达准噶尔小胸鳖甲抗冻蛋白MpAFP698。方法:PCR扩增出mpafp698序列,将其克隆入本室保存的真核表达载体pEGFP-N1中,转染293T细胞;利用RT-PCR、流式细胞仪、免疫荧光、Western印迹检测蛋白表达。结果:构建了真核表达载体pEGFP-N1-mpafp698,在转染后24 h可检测到绿色荧光的表达,而对照组则没有检测到荧光蛋白;Western印迹结果显示表达蛋白的相对分子质量约37 000。结论:为准噶尔小胸鳖甲抗冻蛋白的细胞表达及功能研究奠定了基础。     

High-mobility-group family genes from Lampetra japonica reveal their early origin and molecular evolution in the vertebrate lineage

High-mobility group family (HMG) genes are ubiquitous in vertebrates, including mammals, birds, amphibians and fishes. To elucidate the molecular phylogeny of the HMG genes in the primitive vertebrate, we have cloned three homologues of HMG-box genes, called Lj-HMGB1, Lj-HMGB2 and Lj-HMGBX, from a cDNA library generated from lymphocyte-like cells of the Japanese lamprey (Lampetra japonica), an Agnathan that occupies a critical phylogenetic position between invertebrates and vertebrates. The open reading frames of Lj-HMGB1, Lj-HMGB2 and Lj-HMGBX contained 627 bp, 585 bp and 678 bp, respectively. The analysis of the deduced amino acid sequences indicated that these three putative Lj-HMGB proteins contain four domains: HMG-box A, HMG-box B, an acidic carboxyl-terminal tail and a linker. A phylogenetic analysis revealed that the Lj-HMGB proteins fall outside the vertebrate clade; Lj-HMGBX is descended from a gene ancestral to the mammalian HMGB1/2/3. This discovery implies that there was a gene duplication event in the HMGB1/2/3 gene family that occurred after the divergence of the vertebrates (Cyclostomata) from the Cephalochordata and Urochordata at least 450 million years ago (MYA). The Lj-HMGB1, Lj-HMGB2 and Lj-HMGBX genes were detected in most tissues of the lamprey by RT-PCR. Our findings provide insight into the phylogeny of the HMGB genes in vertebrates.     

Environmental assessment of black locust (<Emphasis Type="BoldItalic">Robinia pseudoacacia</Emphasis> L.)-based ethanol as potential transport fuel

Purpose  

Lignocellulosic ethanol has received special research interest, driven by concerns over high fuel prices, security of energy supplies, global climate change as well as the search of opportunities for rural economic development. A well-to-wheel analysis was conducted for ethanol obtained from black locust (Robinia pseudoacacia L.) by means of the life cycle assessment (LCA) methodology. This study assesses the environmental profile of using ethanol in mixtures E10 and E85 as transport fuel in comparison with conventional gasoline (CG). In addition, the best model of black locust cultivation was analysed under an environmental point of view.     

Isolation and characterization of a <Emphasis Type="Italic">Bacillus licheniformis</Emphasis> strain capable of degrading zearalenone

The worldwide contamination of cereals, oilseeds, and other crops by mycotoxin-producing moulds is a significant problem. Mycotoxins have adverse effects on humans and animals that result in illnesses and economic losses. Reduction or elimination of mycotoxin contamination in food and feed is an important issue. This study aimed to screen soil bacteria for degradation of zearalenone (ZEN). A pure culture of strain CK1 isolated from soil samples showed most capable of degradation of ZEN. Using physiological, biochemical, and 16S rRNA gene sequence analysis methods, CK1 was identified as Bacillus licheniformis. Addition of 2 ppm of ZEN in Luria–Bertani (LB) medium, B. licheniformis CK1 decreased 95.8% of ZEN after 36 h of incubation. In ZEN-contaminated corn meal medium, B. licheniformis CK1 decreased more than 98% of ZEN after 36 h of incubation. In addition, B. licheniformis CK1 was non-hemolytic, non-enterotoxin producing, and displayed high levels of extracellular xylanase, cellulase, and protease activities. These findings suggest that B. licheniformis CK1 could be used to reduce the concentrations of ZEN and improve the digestibility of nutrients in feedstuffs simultaneously.     

Isolation and characterization of an antimicrobial peptide from bovine hemoglobin ��-subunit

In this study, a novel 18-residue linear antimicrobial peptide derived from the central part of the bovine hemoglobin ??-subunit was identified. The peptide was purified by a combination of cationic exchange and reversed-phase high-performance liquid chromatography. The sequence was determined to be VNFKLLSHSLLVTLASHL. The theoretical molecular weight of this peptide was calculated to be 1992.38 Da, which is the same as that determined (1992.401 Da) by matrix-assisted laser desorption ionization mass spectrometry. Sequence analysis showed that there is a high degree of homology in this peptide among hemoglobin ??-subunits of bovine, sheep, deer, porcine, and human. In a radial-diffusion plate assay, this purified peptide exhibited antimicrobial activity against Escherichia coli, Staphylococcus aureus, and Candida albicans.     

中国小赫甲螨属一新种记述(甲螨亚目,小赫甲螨科)(英文)

记述了采自我国西藏和甘肃的小赫甲螨属1新种,郑氏小赫甲螨Hermannniella zhengisp.nov.。模式标本保存在中国科学院动物研究所国家动物博物馆。郑氏小赫甲螨,新种Hermanniella zhengisp.nov.(图1~16)新种以后背板背面小孔状花纹与杜氏小赫甲螨H.dubininiSitnikova,1973、微毛小赫甲螨H.microsetosa Hammer,1966、长毛小赫甲螨H.longisetosaHammer,1966相似,但新种以吻端有凹陷和后背板第3若螨毛f1指向前而不同于这3个已知种。除此之外,新种与杜氏小赫甲螨的区别为:新种梁毛与梁间毛端部尖,感器端部无膨大,被稀疏小刺,后背板背面小孔状花纹的6个点(有时为5或7个)间由细线连接成规则多边形,后缘成体毛4对,光滑;杜氏小赫甲螨梁毛和梁间毛端部钝圆,感器端部被小刺且略膨大,后背板背面花纹为无细线相连的不规则分散小孔,后缘5对成体毛,略被小刺。新种与微毛小赫甲螨的区别为:新种吻毛外表面被小刺,后背板背面花纹为大小均一的小孔,小孔之间有细线连成规则多边形;微毛小赫甲螨吻毛光滑,后背板上的小孔状花纹排列规则,成行但不连接成...     

新疆爱丽缨小蜂属两新种及一新纪录种记述(膜翅目,缨小蜂科)(英文)

记述采自新疆的爱丽缨小蜂属ErythmelusEnock2新种,温泉爱丽缨小蜂Erythmelus(Erythmelus)wenquanensis sp.nov.,短索爱丽缨小蜂Erythmelus(Erythmelus)brevifuniculatus sp.nov.,中国1新纪录种,盲蝽爱丽缨小蜂Erythmelus(Erythmelus)lygivorus Viggianiet Jesu,提供了新疆爱丽缨小蜂属5个种的检索表。模式标本保存在新疆大学生命科学与技术学院昆虫研究室和福建农林大学生物防治研究所。温泉爱丽缨小蜂,新种Erythmelus(Erythmelus)wenquanensis sp.nov.(图1~4)正模♀,新疆温泉县,2001-07-16,胡红英采。副模:2♀♀,来源同正模;1♀,和硕县,2001-08-07,胡红英采;5♀♀,和静县,2001-08-09,胡红英采;2♀♀,托里县,2007-07-29,张铁钢采;1♀,塔城,2007-07-30,张晶晶采;1♀,托里县,2007-07-29,何茜采;1♂,和丰县,2007-08-01,徐建华采。分布:新疆(和硕县、和静县、塔城、和丰县、托里县、温泉县)。新种外部形态特征与E.goochiEnock很相似,主要区别是:新种的第6索节略短于第4、5索节之和;棒节短于柄节至第4索节长度之和;前翅缘毛明显长于翅宽。词源:新种以模式产地命名。短索爱丽缨小蜂,新种Erythmelus(Erythmelus)brevifuniculatussp.nov.(图5~8)正模♀,新疆精河县,2001-07-15,胡红英采。副模:1♀,1♂,来源同正模;6♀♀,4♂♂,乌鲁木齐市,2001-08-22,胡红英采;1♀,新源县,1997-08-07,马德英采;2♀♀,2♂♂,新源县,2001-07-16,胡红英采;1♀,奎屯市,2001-07-17,胡红英采;2♀♀,石河子市,2001-07-12,胡红英采;2♀♀,乌鲁木齐市郊,2001-07-22,孙涛采;1♀,和硕县,2001-08-07,胡红英采;1♀,阿克苏市,2001-06-18,吴卫采;2♀♀,托里县,2007-07-29,徐建华采;1♀,额敏县,2007-07-29,张铁钢采;1♀,额敏县,2007-07-30,何茜采。分布:新疆(阿克苏市、和硕县、乌鲁木齐市、石河子市、奎屯市、精河县、托里县、额敏县、温泉县、新源县)。新种与本属中的任何种都有明显的差异,与另1新种温泉爱丽缨小蜂Erythmelus(Erythmelus)wenquanensis sp.nov.较相似,区别之处:中胸盾片端部淡黄褐色;第4索节等长于第6索节的一半;前翅缘毛长于翅宽的2倍;前翅的长宽比值大于5;雄性外生殖器的长度等长于后足胫节。词源:新种种名brevifuniculatus为组合词,由拉丁词brevi-(意为"短的")和funiculus(意为"索节")组成,指新种索节粗短。盲蝽爱丽缨小蜂Erythmelus(Erythmelus)lygivorus VigganisetJesu中国新纪录(图9~11)研究标本:1♀,新疆托里县,2007-07-29,张铁钢采;1♀,额敏县,2007-07-29,徐建华采。分布:新疆(托里县、额敏县);法国,匈牙利,意大利,西班牙。     

微卫星标记在不同壳色虾夷扇贝家系亲权鉴定的适用性

实验选取8个多态性微卫星位点,用于虾夷扇贝4个不同壳色全同胞和半同胞家系160个子代的亲权鉴定。在亲本未知和一亲本已知的情况下,8个微卫星位点累积排除概率分别为0.823和0.961。鉴于亲权分析时子代的亲本在已知和未知情况下位点的累积排除概率不同,实验采用了两种方法用于家系的亲权鉴定。方法1:当子代的父母本情况未知时,根据子代基因型数据,通过CERVUS 2.0软件计算子代所对应的候选亲本的LOD值,直接将候选亲本中具有最大的两个LOD值的亲本确定为子代的父母本;方法2:在子代的亲本未知情况下,视具有最大LOD值的候选亲本为子代的第一候选亲本,然后将该亲本视为已知,通过CERVUS 2.0软件重新计算每个侯选亲本的LOD值,再从中选择具有最大LOD值的候选亲本作为该子代的第二候选亲本。结果表明,采用方法2得到的家系亲权鉴定成功率达到95%以上,确定了微卫星标记在虾夷扇贝家系鉴定中的可行性。所检测的微卫星位点在子代中出现无效等位基因现象,而无效等位基因存在会引起子代与亲本的错配。实验在家系鉴定时采用了无效等位基因存在(情况1)和缺失(情况2)两种子代基因型文件进行分析,不同情况下同一方法家系鉴定成功率相差无几。这表明了基于多个微卫星位点计算候选亲本LOD值大小寻找子代真实父母本可以降低由无效等位基因引起的错配的几率。研究表明了微卫星标记适合于不同壳色虾夷扇贝家系亲权鉴定工作。