Avian influenza virus infection induces differential expression of genes in chicken kidney

The pathogenic process of highly pathogenic avian influenza virus (HPAIV) infection is poorly understood. To explore the differential expression of kidney genes as a result of HPAIV infection, two cDNA libraries were constructed from uninfected and infected kidneys by suppression subtractive hybridization (SSH). Fifteen genes including IFN-stimulated genes (ISG12), lymphocyte antigen 6 complex locus E gene (LY6E), matrix Gla protein gene (MGP), lysozyme gene, haemopoiesis related membrane protein 1 gene, KIAA1259, MGC68696, G6pc-prov protein gene (G6PC), MGC4504, alcohol dehydrogenase gene (ADH), glutathione S-transferase gene (GST), sodium-dependent high-affinity dicarboxylate transporter gene (SDCT), Synaptotagmin XV (SytXV) and two novel genes were found significantly up-regulated or dramatically suppressed. Differential expression of these genes was further identified by Northern blot. Functional analysis indicated that the regulation of their expression might contribute to the pathogenic process of HPAIV infection. In contrast, the increased expression of three IFN-stimulated genes named ISG12, LY6E, and haemopoiesis related membrane protein 1 gene might reflect host defense responses. Further study showed that ISG12 protein failed to directly interact with NS1 protein of HPAIV which expressed simultaneously in the organs where HPAIV replication occurred, by use of BacterioMatch two-hybrid system. Therefore, our findings may provide new insights into understanding the molecular mechanism underlying the pathophysiological process of HPAIV infection in chicken.     

Response of advance lodgepole pine regeneration to overstory removal in eastern Idaho

Twenty-two stands of advance lodgepole pine released with overstory removal were sampled to determine height growth response. Tree and site characteristics correlated with release response were identified, and a mathematical model was developed to predict height growth in years 6 through 10 after release as a function of residual overstory basal area, height at release, percent rock cover, five-year cumulative growth prior to release, logging damage, stand elevation, and habitat type. Lodgepole pine responded to release with increased height growth in 97% of the trees sampled. Growth was similar to that of unsuppressed trees. Height growth is best when the entire overstory is removed and logging damage is avoided. Taller trees generally do not respond as well as shorter trees. However, trees growing fast before harvest continue to grow fast regardless of their height at release. Recommendations for selecting stands of advance lodgepole pine to release are provided.     

Herbicide-tolerant crops—real farmer opportunity or potential environmental problem?

Classical breeding, in-vitro selection and genetic engineering techniques have produced herbicide-tolerant crops. Commercial adoption of these cultivars in North America provides tolerance predominantly to non-selective herbicides for novel weed-control strategies. Of special value is the integration of these crops in minimum tillage situations, maintaining of a wide range of herbicides with different modes of action to provide a variety of opportunities for weed-control management. Associated with these special crops are a series of environmental issues which at present limit the rate of commercial development in Europe. Unlike the successful performance of herbicide tolerance in the crops, these strategic issues are much more difficult to resolve for technical, political, ethical and moral reasons. The primary concerns are the feasibility of controlling the volunteer crop and the opportunity for indiscriminate introgression of the herbicide-tolerance gene into agricultural and natural ecosystems. It is unlikely that these questions will be resolved without scaling-up field experiments to include the detection of herbicide-tolerance genes. The economic and management implications of herbicide-tolerant crops require special consideration in view of the necessity to integrate conventional and transgenic crops in new cropping systems. © 1998 SCI     

甘蓝型油菜种子不同发育期基因差异表达cDNA文库构建

以授粉后27 d的种子mRNA为目标群体,授粉后7 d的种子mRNA为对照,构建甘蓝型油菜种子发育期基因差异表达文库.文库包括560个克隆,经菌落PCR检测,456个菌落含有插入片段,占全部克隆的91.2 %,片段大小200～600 bp.对PCR阳性克隆进行斑点杂交,得到201个上调的阳性斑点杂交阳性克隆,对其进行测序,得到有效ESTs序列198个.BLASTN结果显示,134个ESTs序列没有同源序列,可能是新基因,另外64个ESTs序列在油菜或拟南芥核酸数据库中存在同源序列.这64个ESTs序列和已知功能(主要是能量代谢、物质代谢、基因调控、衰老及抗性等)的蛋白有高度相似性.     

应用抑制性差减杂交技术构建副猪嗜血杆菌基因组差减文库

为构建副猪嗜血杆菌(Haemophilus parasuis,HPS)有毒力菌株SW124(血清4型)和无毒力菌株H465(血清11型)间的差异表达基因文库,采用抑制性差减杂交技术(Suppression subtractive hybridization,SSH)分析SW124和H465菌株细菌基因组的表达差异,并进行两轮差减杂交和两次PCR扩增,将第2次PCR产物与pMD19-T载体相连,转化大肠杆菌E.coli DH5α感受态细胞并进行文库扩增和蓝白斑筛选,RT-PCR鉴定差异表达文库。结果共获得327个阳性克隆,筛选100个克隆制备质粒进行PCR检测,均扩增出100～2 000 bp大小的片段。差异DNA差减文库的构建为进一步筛选、克隆HPS特异的未知新基因、建立分子鉴别新体系奠定了基础。     

枯萎病菌诱导的节瓜抑制差减cDNA文库的构建

以抗枯萎病节瓜品种杂交20号为材料,利用抑制差减杂交(Suppression subtractive hybridization,SSH)技术,构建了枯萎菌(Fusarium oxysporum f.sp.beniacasa)诱导的节瓜SSH-cDNA文库。文库质量检测表明,差减杂交效率较高,质量较好。得到正向差减文库的滴度为3.64×105 CFU/mL,反向差减文库滴度为6.80×105 CFU/mL;正向和反向差减文库的重组率均大于90%,插入片段在400～800bp之间。     

凡纳滨对虾肌球蛋白轻链基因MLC的克隆及表达分析

以生长性状分离凡纳滨对虾家系的肌肉组织,构建凡纳滨对虾生长性状差减cDNA文库,斑点杂交筛选获得MLC基因,克隆获得全长编码区序列.序列分析表明序列包含462 bp的开放阅读框,推测编码的蛋白质为153氨基酸,预测的分子量为17.5ku,等电点为4..67.通过荧光定量PCR的方法研究了MLC基因在凡纳滨对虾不同组织和不同生长期的肌肉组织中表达情况,显示MLC基因在凡纳滨对虾的心脏、肝胰腺、肌肉、胃、肠、眼柄、鳃和血等组织或器官中都有表达,在肌肉组织中表达量最高,在肝胰腺中表达量最低;在日龄为45d的对虾肌肉组织中表达量最高,在日龄为10 d的对虾肌肉组织中表达量最低.     

白桦雌花发育前期和中期差异基因表达分析

白桦为雌雄同株的单性花树种,在木本植物花发育研究中有着重要作用。本文旨在初步确定白桦雌花发育过程中的关键时期大孢子发生(前期)和配子体发育(中期)过程中基因表达及调控的差异,同时挖掘与雌花发育相关的基因。首先,运用细胞学方法确定试材雌花的发育时期。其次,采用抑制消减杂交(SSH)技术,分别以白桦前期和中期雌花序的cDNA作为tester和driver,建立正、反向抑制消减杂交cDNA文库:BHHQ和BHHZ。共获得1406个高质量的EST序列,其中787个与数据库中的已知核酸或蛋白序列有较高同源性, 47个可能与花发育相关。GO分析结果表明,这些基因34.96%参与生物合成,16.45%参与细胞组件,48.59%参与分子功能。另外,选择6个与花发育相关EST进行qRT-PCR分析,结果显示这些基因在不同组织中都有一定表达。其中bhhq.408,bhhq.435,bhhz.359,bhhz.215在生殖器官中的表达明显高于营养器官;bhhq.254,bhhq.190在绝大多数组织中均有一定表达,说明这些基因不仅参与生殖器官的发育,也在营养器官中发挥一定的作用。