草菇低温胁迫初期应答关键通路的探究

本研究利用表达谱芯片技术解析了低温胁迫初期（0-120min）不同低温处理时间草菇转录组水平的变化。芯片结果分析发现在低温诱导过程中,低温处理20min内变化基因的功能为单加氧酶、氧化还原酶和细胞内的氧化还原。低温处理40min时表达差异基因具有结合RNA的功能。80min的时间段内,基因的功能比较集中于核酸物质和能量的代谢。CYP450的代谢途径在20-40min、60-80min和100-120min低温处理的菌丝体中均有显著性变化,表明这一通路在草菇低温处理过程中表现活跃。基因表达趋势分析发现34个表达差异基因富集到显著表达趋势模型,基因共表达网络分析发现VVO_04066位于网络的核心地位,推测VVO_04066通过提高磷酸肌醇特异性磷脂酶C（PI-PLC）的胞内水平,促进糖蛋白和几丁质的合成,从而完成草菇细胞的低温胁迫应答。     

极端耐盐碱菌株的筛选及其菌肥对盐碱条件下小麦生长和土壤环境的影响

为获得具有盐碱地改良应用潜力的耐盐碱菌株,将采集于东营盐碱地的土样稀释涂布于pH 9、盐浓度100 g·L-1的Gibbson改良培养基上,共获得18株细菌.通过提高盐浓度、pH值,最终获得在pH 12、盐浓度20%的条件下仍然可以生长的极端耐盐碱菌株N14.对N14进行形态学、生理生化特征和16S rDNA序列分析鉴定,结果表明:菌株N14为马氏芽孢杆菌.盆栽试验结果表明,与盐碱土(CK)相比,N14菌肥可以显著提高小麦的生物量,株高、鲜重、干重分别提高了21.8%、57.9%、41.7%;显著增加小麦叶绿素a、叶绿素b、叶绿素总量,增长率分别为36.4%、20.0%、31.7%;显著提高盐碱土壤中的蔗糖酶、脲酶和碱性磷酸酶的活性,增长率分别为23.2%、68.8%、106.5%;显著提高小麦根系的超氧化物歧化酶、过氧化物酶和过氧化氢酶的活性,增长率分别为109.6%、17.8%、50%;显著减少小麦根系丙二醛的含量,减少率为39.8%.本研究为极端耐盐碱菌的应用提供了一条新思路,为盐碱地的改良提供了一条新途径.     

黑麂线粒体基因组序列分析

采用PCR产物直接测序方法测定了黑麂线粒体基因组全序列 ,初步分析了其基因组特点并定位了各基因的位置 .结果显示 :黑麂的线粒体基因组全序列长度为 1 6 35 7bp ,可编码 2 2种tRNA、2种rRNA、1 3种蛋白质 ,碱基组成及基因位置与小麂、赤麂和其它哺乳类动物的线粒体基因组相似 ;模拟电子酶切图谱与先前的报道基本一致 ;基于细胞色素b的全基因序列 ,分别以最大简约法、N J法、最大似然数法与其它 1 4种鹿类动物的相应序列进行了聚类分析 ,构建出相似的系统进化树 :初步确定了麂亚科动物在鹿科中处于与鹿亚科、北美鹿亚科并列的进化地位 .在此基础上 ,进一步以黑麂、赤麂、小麂的线粒体编码RNA和编码蛋白质的基因序列构建系统进化树 ,分析了三者的亲缘关系 .结果表明 :黑麂和赤麂亲缘关系较近 ,是较新的物种 ,而小麂是较为原始的物种     

Risks of Avian Influenza Transmission in Areas of Intensive Free-Ranging Duck Production with Wild Waterfowl

For decades, southern China has been considered to be an important source for emerging influenza viruses since key hosts live together in high densities in areas with intensive agriculture. However, the underlying conditions of emergence and spread of avian influenza viruses (AIV) have not been studied in detail, particularly the complex spatiotemporal interplay of viral transmission between wild and domestic ducks, two major actors of AIV epidemiology. In this synthesis, we examine the risks of avian influenza spread in Poyang Lake, an area of intensive free-ranging duck production and large numbers of wild waterfowl. Our synthesis shows that farming of free-grazing domestic ducks is intensive in this area and synchronized with wild duck migration. The presence of juvenile domestic ducks in harvested paddy fields prior to the arrival and departure of migrant ducks in the same fields may amplify the risk of AIV circulation and facilitate the transmission between wild and domestic populations. We provide evidence associating wild ducks migration with the spread of H5N1 in the spring of 2008 from southern China to South Korea, Russia, and Japan, supported by documented wild duck movements and phylogenetic analyses of highly pathogenic avian influenza H5N1 sequences. We suggest that prevention measures based on a modification of agricultural practices may be implemented in these areas to reduce the intensity of AIV transmission between wild and domestic ducks. This would require involving all local stakeholders to discuss feasible and acceptable solutions.     

二元融合子与荧光假单胞菌耐药性遗传标记的筛选

应用定性和定量药物敏感性试验与诱变相结合,筛选出荧光假单胞菌56-12-10菌株的遗传标记为链霉素抗性,二元融合子AM-1菌株的遗传标记为环丙沙星抗性,确定了在这两种细菌原生质体融合试验中,选择培养基中链霉素的应用浓度为300Iu/ml,环丙沙星应用浓度为100Iu/ml。     

RT—PCR法检测大鼠脑组织及C6细胞中β—APP的表达

β-淀粉样蛋白(β-AP)是阿尔茨海默氏病(Alzheimer’s disease)病人老年斑的主要成分,它是β-淀粉样前体蛋白(β-APP)剪切后的产物。β-APP基因在体内存在β-APP_(695),β-APP_(751)β-APP_(770)。等几种主要的转录物,它们的区别在于Kunize丝氨酸蛋白酶抑制区(KDI)编码序列的存在和缺失。通过RT-PCR技术,用针对KPI编码区两侧序列的一种特异性引物可由总RNA样品中扩增出反映以上三种转录物的cDNA片段。实验表明,胎鼠脑组织中未检测到β-APPmRNA;6月龄大鼠海马组织,大脑皮层中只检测到β-APP_(695);在神经胶质瘤细胞系C6中存在β-APP_(695),β-APP_(751),β-APP_(770),其中β-APP_(770)占优势。     

Nuclear targeting signal recognition: a key control point in nuclear transport?

Recent progress indicates that there are multiple pathways of nucleocytoplasmic transport which involve specific targeting sequences, such as nuclear localization sequences (NLSs), and cytosolic receptor molecules of the importin/karyopherin superfamily which recognise and dock the NLS-containing proteins at the nuclear pore. This first step of nuclear import/export is of central importance, with the affinity of the importin-targeting sequence interaction a critical parameter in determining transport efficiency. Different importins possess distinct NLS-binding specificities, which allows the system to be modulated through differential expression of the importins themselves, as well as through competition between different importins for the same protein, and between different proteins for the same importin. The targeting sequence-importin interaction can also be influenced directly by phosphorylation increasing the affinity of the interaction with importins or by targeting sequence masking through phosphorylation or specific protein binding. Targeting sequence recognition thus appears to represent a key control point in the regulation of nuclear transport. BioEssays 22:532-544, 2000.     

Blockade of L-type voltage-gated Ca channel inhibits ischemia-induced neurogenesis by down-regulating iNOS expression in adult mouse

Neurogenesis in the adult mammalian hippocampus may contribute to repairing the brain after injury. The signals that regulate neurogenesis in the dentate gyrus following ischemic stroke insult are not well known. We have previously reported that inducible nitric oxide synthase (iNOS) expression is necessary for ischemia-stimulated neurogenesis in the adult dentate gyrus. Here, we show that mice subjected to 90 min of middle cerebral artery occlusion (MCAO) significantly increased the number of new neurons and up-regulated iNOS expression in the dentate gyrus. Blockade of the L-type voltage-gated Ca(2+) channel (L-VGCC) prevented neurogenesis in the dentate gyrus and subventricular zone (SVZ), and down-regulated iNOS expression in the dentate gyrus after cerebral ischemia. This study suggests that Ca(2+) influx through L-VGCC is involved in ischemia-induced neurogenesis by up-regulating iNOS expression.     

Kinetic FP-TDI assay for SNP allele frequency determination

Strategies for identifying genetic risk factors in complex diseases by association studies require the comparison of allele frequencies of numerous SNPs between affected and control populations. Theoretically, hundreds of thousands of SNP markers across the genome will have to be genotyped in these studies. Genotyping SNPs one sample at a time is extremely costly and time consuming. To streamline whole genome association studies, some have proposed to screen SNPs by pooling the DNA samples initially for allele frequency determination and perform individual genotyping only when there is a significant discrepancy in allele frequencies between the affected and control populations. Here we describe a new method for determining the allele frequency of SNPs in pooled DNA samples using a two-color primer extension assay with real-time monitoring of fluorescence polarization (named kinetic FP-TDI assay). By comparing the ratio of the rate of incorporation of the two allele-specific dye-terminators, one can calculate the relative amounts of each allele in the pooled sample. The accuracy of allele frequency determination with pooled samples is within 3.3 +/- 0.8% of that determined by genotyping individual samples that make up the pool.