肠出血性大肠杆菌毒力因子Z1444的原核表达及其丝/苏氨酸激酶活性验证

目的:在大肠杆菌中表达肠出血性大肠杆菌(EHEC)毒力岛上的毒力因子Z1444并纯化,对其丝/苏氨酸激酶活性进行初步检测。方法:根据GenBank中Z1444基因序列及pET-28a(+)载体的多克隆位点设计引物,以EHECO157∶H7全菌裂解液为模板,经PCR钓取1047 bp的目的片段,与表达载体pET-28a(+)连接,构建重组表达质粒pET-28a(+)-Z1444,将其转化至大肠杆菌BL21(DE3)中,IPTG诱导蛋白表达并经SDS-PAGE鉴定,利用体外反应体系鉴定重组蛋白的丝/苏氨酸激酶活性。结果:双酶切和测序鉴定表明,pET-28a(+)-Z1444原核表达质粒构建正确;诱导表达后经纯化,获得纯度在90%以上的可溶性重组Z1444,相对分子量约为38×103;体外酶活实验验证了Z1444的丝/苏氨酸激酶活性。结论:Z1444在大肠杆菌中获得高效可溶性表达,为后续功能验证奠定了基础。     

固相微萃取结合GC/MS联用技术研究野松茸干品的挥发性成分

采用顶空固相微萃取结合气相色谱-质谱联用技术分析鉴定了野松茸干品中的挥发性风味成分.鉴定出48种风味化合物,占挥发性成分总量的91.44％,其中酸类9种、酯类6种、含氮杂环化合物11种、醇类3种、醛类5种、酮类10种、其他类4种.野松茸干品中主要的芳香成分是桂酸甲酯(42.99％)、3-甲基丁酸(8.56％)、2,6-二甲基吡嗪(8.36％)、2,5-二甲基吡嗪(2.46％)和1-辛烯-3-醇(2.06％).     

台湾山柚属(山柚子科)的一个新组合

木文是对《甜菜树属(中国山柚子科新记录属)一新种》(载“云南植物研究”1989年11卷4期)一文的修订。将误定于甜菜树属(Mlelientha Pierre)下的长蕊甜菜树(M.longi-staminea)改隶于台湾山柚属(Champereia Griff.),重新组合为四数台湾山柚(Champereia)longistaminea。如果将长蕊甜菜树(Melientha longistaminea)置于甜菜树属(Melientha)中,则混淆了台湾山柚属(Champereia Pierre)与甜菜树属(Melientha Pierre)的区别界线。     

Insecticidal activity of the chitinase from the Spodoptera litura nucleopolyhedrovirus

Baculovirus chitinase gene (chiA) is a late gene essential for liquefying the host insect at a late stage of infection for its hydrolyzing chitin function. In a previous report, baculovirus ChiA has been shown to offer many interesting new opportunities for pest control. Recently, a putative chiA gene was identified in the Korean isolate of the Spodoptera litura nucleopolyhedorvirus (SpliMNPV‐K1) genome. The open reading frame (ORF) contains 1692 nucelotides and encodes a protein of 563 amino acids with a predicted molecular weight of about 62.6 kDa. To study the insecticidal activity of ChiA from SpliMNPV‐K1, we constructed a recombinant AcMNPV, Ap‐SlChiA, which is designed to express the ChiA under the control of a polyhedrin promoter. Western blot analysis indicated that ChiA was successfully expressed by this recombinant virus. Chitinase assay revealed that the chitobiosidase and endochitinase activity of the recombinant virus was 2.5‐ and 3.9‐flods higher than those of wild‐type AcMNPV, respectively. In addition, the recombinant virus showed higher evident insecticidal activity against 3rd instar larvae of Spodotera exigua than that of the AcMNPV. These results suggest that the chiA gene from SpliMNPV‐K1 could be successfully applied to improve pathogenicity of baculoviruses.     

褐飞虱抗药性研究现状

褐飞虱Nilaparvata lugens(Stal)对杀虫剂产生抗药性是其近年来暴发频繁的重要原因。文章综述国内外关于褐飞虱抗药性的研究成果,包括褐飞虱抗性测定方法、抗药性的发展、交互抗性、抗性遗传、抗性机理及抗性治理等。田间褐飞虱种群对新烟碱类药剂产生不同程度的抗药性,其中对吡虫啉产生高水平到极高水平抗性,对氯噻啉和噻虫嗪分别产生中等水平和低水平的抗药性,对呋虫胺和烯啶虫胺仍然处于敏感性阶段。此外,褐飞虱种群对噻嗪酮(昆虫生长调节剂)产生低水平到中等水平抗性。长期大面积使用化学药剂是褐飞虱产生抗药性的重要原因。因此,必须加强褐飞虱的抗性治理,以延缓其抗药性进一步发展。     

Properties of STAT1 and IRF1 enhancers and the influence of SNPs

Background

STAT1 and IRF1 collaborate to induce interferon-γ (IFNγ) stimulated genes (ISGs), but the extent to which they act alone or together is unclear. The effect of single nucleotide polymorphisms (SNPs) on in vivo binding is also largely unknown.

Results

We show that IRF1 binds at proximal or distant ISG sites twice as often as STAT1, increasing to sixfold at the MHC class I locus. STAT1 almost always bound with IRF1, while most IRF1 binding events were isolated. Dual binding sites at remote or proximal enhancers distinguished ISGs that were responsive to IFNγ versus cell-specific resistant ISGs, which showed fewer and mainly single binding events. Surprisingly, inducibility in one cell type predicted ISG-responsiveness in other cells. Several dbSNPs overlapped with STAT1 and IRF1 binding motifs, and we developed methodology to rapidly assess their effects. We show that in silico prediction of SNP effects accurately reflects altered binding both in vitro and in vivo.

Conclusions

These data reveal broad cooperation between STAT1 and IRF1, explain cell type specific differences in ISG-responsiveness, and identify genetic variants that may participate in the pathogenesis of immune disorders.

     

Anti-apoptotic and Anti-oxidative Roles of Quercetin After Traumatic Brain Injury

Experimental studies have demonstrated significant secondary damage (including cell apoptosis, blood–brain barrier disruption, inflammatory responses, excitotoxic damage, and free radical production) after traumatic brain injury (TBI). Quercetin is a natural flavonoid found in high quantities in fruits and vegetables, and may be a potential antioxidant and free radical scavenger. The purpose of this study was to determine the effects of quercetin on TBI-induced upregulation of oxidative stress, inflammation, and apoptosis in adult Sprague–Dawley rats. Animals were subjected to Feeney’s weight-drop injury, thus inducing the parietal contusion brain injury model. Quercetin was administered (30 mg/kg intraperitoneal injection) 0, 24, 48, and 72 h after TBI. Quercetin reduced cognitive deficits, the number of TUNEL- and ED-1-positive cells, the protein expressions of Bax and cleaved-caspase-3 proteins, and the levels of TBARS and proinflammatory cytokines, and increased the activity of antioxidant enzymes (GSH-Px, SOD, and CAT) at 1 week after TBI. Our results suggest that in TBI rats, quercetin improves cognitive function owing to its neuroprotective action via the inhibition of oxidative stress, leading to a reduced inflammatory response, thereby reducing neuronal death.     

云南食用菌眼蕈蚊分类及优势种分析

本文报道了云南省各食用菌栽培地眼蕈蚊科（Sciaridae）害虫共6属11种, 其中异迟眼蕈蚊Bradysia difformis Frey为中国新纪录种, 独刺普眼蕈蚊Cosmosciara perniciosa Edwards为中国新记录属和种, 并提供了云南省食用菌眼蕈蚊种类检索表。研究标本均保存于云南农业大学昆虫系标本室。根据各地标本采集数据对不同食用菌上眼蕈蚊种类优势度进行了分析, 结果表明平菇厉眼蕈蚊Lycoriella pleuroti Yang et Zhang和异迟眼蕈蚊B. difformis Frey为云南省食用菌上眼蕈蚊的主要优势种类。     

有机配体对稀土元素在小麦体内积累和分异的影响

采用营养液培养和添加外源混合稀土等方法,研究了有机配体柠檬酸、EDTA和DTPA对稀土元素在小麦的根和叶中积累与分异的影响。结果表明,低浓度有机配体对小麦根和叶中的稀土元素,尤其是轻稀土元素的积累有轻微的促进作用,随浓度的升高则表现出显著的降低作用。有机配体对重稀土的作用比轻稀土强,使根和叶中稀土元素的分布曲线向重稀土相对亏缺的方向发展。3种配体对轻、重稀土分异的作用强度为:EDTA>DTPA>柠檬酸。通过VM INTEQ计算表明,在EDTA和DTPA作用下小麦叶中稀土元素的积累与轻、重稀土的分异主要由溶液中呈自由离子态稀土元素的含量和组成控制;柠檬酸作用下小麦叶中稀土元素的变化与自由离子态稀土的含量和组成关系较弱,推测REE-柠檬酸络合物可被小麦直接吸收并运转到小麦的叶中。