Scytovirin在大肠杆菌中的可溶性表达

目的:获得Scytovirin(SVN)蛋白及其多克隆抗体.方法:按照NCBI上公布的SVN基因序列设计合成引物,合成SVN基因,构建pET32c-SVN原核表达重组质粒,经限制性酶切分析、DNA序列测定插入片段正确;将该重组质粒转化大肠杆菌BL21(DE3),IPTG诱导重组蛋白表达;用离子交换层析法及金属亲合层析法纯化蛋白,采用Tris-Tricine系统分析;以经过纯化的蛋白为抗原免疫白兔,制备SVN多克隆抗体.结果:对表达产物进行了分离纯化,SVN纯度达到91%;用纯化的样品制备了多克隆抗体,抗血清效价为1∶102 400.结论:SVN在大肠杆菌表达系统中获得了高效可溶表达,并制备了其多克隆抗体,为进一步深入研究SVN提供了材料.     

HPV转基因动物研究进展

人乳头状瘤病毒(Human papillomavirus,HPV)可以引起人宫颈癌、肛门生殖器癌、尖锐湿疣等多种良、恶性肿瘤,严重威胁人类健康。然而,由于HPV严格的嗜人类上皮组织的特性,自然状态下不感染任何实验动物,给研究HPV的感染过程、致病机理,以及开发治疗HPV感染的药物等带来了困难,因此,研究HPV的动物模型显得尤为重要。     

密穗型水稻品种籽粒垩白性状改良研究

采用籽粒长宽比较大、穗部着粒密的散穗型材料(EG23)改良粳稻密穗型品种的籽粒垩白性状.结果表明,经改良后得到的密穗型品系EA6,与原亲本浙粳20比较,其穗部长度缩短,每穗总粒数增加,着粒密度增大,而籽粒垩白特性得到明显的改善,表明在穗部长度和着粒结构未得到改良的情况下,调节籽粒长宽比对改善密穗型品种籽粒垩白性状具有可能性.穗部不同粒位籽粒垩白性状改良的效果不同,穗顶部和穗中部的改良效果明显优于穗基部.设计的4个不同杂交配组方式中,以反回交配组方式(浙粳20/ EG23//浙粳20)选育效果最好.EA6具有较好的农艺性状,既可作为优异种质资源利用,也可直接应用于生产.这一结果从育种实践上较好地协调了密穗型品种高产与优质的矛盾,对于培育既有密穗型的高产株型又有优良籽粒外观品质的水稻品种具有重要意义.     

应用基因芯片技术检测肝组织的基因表达

为检测正常肝组织中的基因表达状况,采用cDNA microarray技术对正常肝组织中表达的1500个基因进行定量,结果为97个基因较高表达,1010个基因中度表达,380个基因低表达,结果是cDNA microarray技术是大规模检测基因表达的有效方法。     

Involvement of metabotropic glutamate receptors in excitatory amino acid and GABA release following spinal cord injury in rat

Spinal cord injury (SCI) leads to an increase in extracellular excitatory amino acid (EAA) concentrations resulting in glutamate receptor-mediated excitotoxic events. The glutamate receptors include ionotropic (iGluRs) and metabotropic (mGluR) receptors. Of the three groups of mGluRs, group-I activation can initiate intracellular pathways that lead to further transmitter release. Groups II and III mGluRs function mainly as autoreceptors to regulate neurotransmitter release. In an effort to examine the role of mGluRs in the increase in EAAs following SCI, we administered AIDA, a potent group-I mGluR antagonist immediately after injury. To determine subtype specific roles of the group-I mGluRs, we evaluated EAA release following LY 367385 (mGluR1 antagonist) and MPEP (mGluR5 antagonist) administration. To evaluate group-II and -III mGluRs we administered APDC (group-II agonist) and L-AP4 (group-III agonist) immediately following injury; additionally, we initiated treatment with CPPG (group-II/-III antagonist) and LY 341495 (group-II antagonist) 5 min prior to injury. Subjects were adult male Sprague-Dawley rats (225-250 g), impact injured at T10 with an NYU impactor (12.5 mm drop). Agents were injected into the epicenter of injury, amino acids where collected by microdialysis fibers inserted 0.5 mm caudal from the edge of the impact region and quantified by HPLC. Treatment with AIDA significantly decreased extracellular EAA and GABA concentrations. MPEP reduced EAA concentrations without affecting GABA. Combining LY 367385 and MPEP resulted in a decrease in EAA and GABA concentrations greater than either agent alone. L-AP4 decreased EAA levels, while treatment with LY 341495 increased EAA levels. These results suggest that mGluRs play an important role in EAA toxicity following SCI.     

RAPD分析在绢丝昆虫亲缘关系研究中的应用Ⅱ.柞蚕品种间的遗传差异

采用RAPD技术,对5个柞蚕品种的遗传差异进行比较研究.结果表明,所采用的40个随机引物中,有27个引物扩增谱带清晰且重复性较好,扩增总片段数253条,单个引物的扩增片段数在4～16之间,片段大小在0.33～3.0kb之间.不同柞蚕品种间的遗传差异较小,遗传距离(D)在0.066～0.1659之间,根据D值,由UPGMA聚类分析软件绘制了它们的分子进化树。 Abstract:Random amplified Polymorphic DNA (RAPD) was used to analyze the genetic diversity among Antheraea pernyi.The genetic variance of five Antheraea pernyi was studied.The result showed that:27 of 40 arbitrary primers could amplify clear and repeating bands.A total of 262 fragments were obtained.Each primer gave 4~16 bands and the average was 9.7.The length of the band was 0.33~3.0kb.The D value between different breeds of Antheraea pernyi was 0.066~0.1659.The D value was used to construct a dendrogram by UPGMA.     

CO2浓度倍增对小麦叶绿体超微结构、超分子结构及光谱特性的影响

超薄切片及冰冻撕裂电镜观察、吸收光谱及 77K低温荧光发射光谱的测定结果表明 :CO2 浓度倍增对小麦(TriticumaestivumL .)叶绿体的超微、超分子结构及光谱特性的影响均为正效应。具体反映在 :(1)小麦叶绿体中除了比对照积累有较多的淀粉粒外 ,其基粒和基质类囊体膜发育较好 ;(2 )叶绿体的光合膜系 ,无论是垛叠和非垛叠膜区 ,其镶嵌于内质膜撕裂面 (EFs和EFu)及原生质膜撕裂面 (PFs和PFu)的功能蛋白粒均比其对照的发育良好 ,尤其PFs与EFs面较为突出 ,即它们除了所含蛋白粒的密度较大外 ,在EFs面上有时还呈现出密集有序的阵列结构 ;(3)叶绿体整个吸收谱带 ,尤其红区和蓝区的主峰均较其对照有较大的光吸收 ,表明对光能的捕获能力明显高于对照 ;(4)无论是以 4 36nm还是以 4 80nm波长激发的 ,其叶绿体的F684/F73 3 (PSⅡ /PSⅠ )的比值均较对照的高 ,表明CO2 浓度倍增条件下生长的小麦叶片叶绿体的PSⅡ相对荧光强度有所增强 ,这与叶绿体的超微、超分子结构及吸收光谱的测定结果相一致。以上结果可为小麦在高CO2 浓度下增产提供理论依据。     

Abundances and host relationships of chigger mites in Yunnan Province,China

This paper reports on ectoparasitic chigger mites found on small mammals in Yunnan Province, southwest China. Data were accumulated from 19 investigation sites (counties) between 2001 and 2009. A total of 10 222 small mammal hosts were captured and identified; these represented 62 species, 34 genera and 11 families in five orders. From the body surfaces of these 10 222 hosts, a total of 92 990 chigger mites were collected and identified microscopically. These represented 224 species, 22 genera and three subfamilies in the family Trombiculidae (Trombidiformes). Small mammals were commonly found to be infested by chigger mites and most host species harboured several species of mite. The species diversity of chigger mites in Yunnan was much higher than diversities reported previously in other provinces of China and in other countries. A single species of rodent, Eothenomys miletus (Rodentia: Cricetidae), carried 111 species of chigger mite, thus demonstrating the highest species diversity and heaviest mite infestation of all recorded hosts. This diversity is exceptional compared with that of other ectoparasites. Of the total 224 mite species, 21 species accounted for 82.2% of all mites counted. Two species acting as major vectors for scrub typhus (tsutsugamushi disease), Leptotrombidium scutellare and Leptotrombidium deliense, were identified as the dominant mite species in this sample. In addition to these two major vectors, 12 potential or suspected vector species were found. Most species of chigger mite had a wide range of hosts and low host specificity. For example, L. scutellare parasitized 30 species of host. The low host specificity of chigger mites may increase their probability of encountering humans, as well as their transmission of scrub typhus among different hosts. Hierarchical clustering analysis showed that similarities between different chigger mite communities on the 18 main species of small mammal host did not accord with the taxonomic affinity of the hosts. This suggests that the distribution of chigger mites may be strongly influenced by the environment in which hosts live.