桑树桑黄菌丝体和子实体基因差异表达分析

通过对桑树桑黄Sanghuangporus sanghuang菌丝体和子实体2个不同生长阶段的转录组进行分析,为研究桑黄子实体生长发育相关机制奠定基础。采用Illumina测序技术,对桑树桑黄菌株S23菌丝体和子实体2个不同生长发育阶段进行了全转录组测序。将转录组测序reads比对到参考序列上,菌丝体测序样本的reads比对率为82.89%;子实体测序样本的reads比对率为83%。基因差异表达分析显示,与菌丝体相比,子实体中显著上调表达基因为2 898个,显著下调表达基因为1 965个。经过Blast nr比对发现,桑黄菌在子实体阶段表达量上升的基因主要与各种氧化酶活性、疏水蛋白等相关;表达量下降的基因主要与糖类、氨基酸结合、运输等相关。基因本体（gene ontology,GO）富集分析表明,菌丝体及子实体两个阶段与跨膜转运相关的差异表达基因富集明显。代谢通路（pathway）富集分析表明,类固醇生物合成、精氨酸生物合成、丝裂原活化蛋白激酶（mitogen-activated protein kinase,MAPK）信号通路等差异基因富集明显。     

五指山猪和长白猪背最长肌差异基因的筛选与注释

为了筛选五指山猪和长白猪背最长肌组织差异表达基因,本研究采用RNA-seq技术和生物信息学方法对6月龄、8月龄五指山猪和长白猪的背最长肌进行转录组测序分析,并对差异表达基因进行GO和KEGG Pathway显著性富集分析。通过与猪基因组比对分析后发现,在6月龄和8月龄五指山猪和长白猪均有约44 000 000条Clean reads能够比对到猪基因组上相关基因,每个文库均获得约18 200个表达基因,而转录本的数据为25 000个。在6月龄五指山猪与长白猪中共有30个基因差异表达,在8月龄五指山猪与长白猪中获得29个差异表达的基因。这些差异表达的基因主要富集在与糖酵解代谢、MAPK信号代谢通路和胰岛素信号通路等肌肉发育信号通路,通过筛选获得了与肌肉生长发育有关的候选基因,为探索五指山猪肌肉发育的分子机制提供了理论依据。     

青藏高原地区3种蒿属植物转录组比较分析

米蒿（Artemisia dalai-lamae）、冷蒿（Artemisia frigida）和臭蒿（Artemisia hedinii）作为西北干旱半干旱地区常见植物,具有防风固沙、阻止草原退化、杀菌消炎和治疗多种疾病的功效。通过Illumina测序技术分别对3种蒿属植物进行转录组测序和分析,分别获得54 268 322,46 434 864和43 971 646条Clean reads,并对测序结果进行了SNP和可变剪接分析。在米蒿和冷蒿对比组（ADL vs AF）中筛选到6 107个差异基因,在米蒿和臭蒿对比组（ADL vs AH）中筛选到4 822个差异基因,在冷蒿和臭蒿对比组（AF vs AH）中筛选到3 755个差异基因。GO富集将差异基因注释到生物过程、细胞组成和分子功能这3个类别共47个条目。KEGG代谢通路分析将3个对比组的差异基因分别富集到198、198和197条通路。对每个对比组的前10个高表达差异基因进行了分析,并筛选出25个生物碱、萜类和类黄酮的活性成分可能相关的差异基因。本研究为蒿属植物的物种鉴定、抗逆性研究和资源利用提供一定的科学依据。     

暗纹东方鲀线粒体COI及其侧翼tRNA基因的克隆与序列分析

以暗纹东方鲀(Takifugu fasciatus)肝脏的线粒体DNA为模板,按照红鳍东方鲀线粒体DNA序列设计合成特异引物进行PCR扩增,克隆并测定了线粒体细胞色素氧化酶I亚基（COI）及其侧翼tRNA基因的全序列,结果显示,克隆了暗纹东方鲀COI基因1546bp及其5′端上游的tRNATyr基因和3′端下游的tRNASer基因序列共1766bp。用DNA分析软件对暗纹东方鲀与GenBank中10个目13种鱼类的COI序列进行比较分析,显示暗纹东方鲀与这些鱼类的COI基因具有较高的同源性,与同属红鳍东方鲀的同源性最高为97.6%,与同目不同科的矛尾翻车鲀和翻车鲀的同源性为76.5%和75.4%。根据暗纹东方鲀与其他13种鱼的COI基因序列同源性所建立的进化树,与传统的分类地位基本吻合。推定的这二种tRNA的二级结构都具有典型的三叶草型结构。     

东方鲀生长激素基因3''''-UTR多态性分析

以暗纹东方鲀（Takifugu. Obscurus）、红鳍东方鲀（Takifugu. Rubripes）、星点东方鲀（Takifugu. niphobles）共82个个体为对象,运用单链构向多态性（SSCP）技术和测序技术分析生长激素（Growth Hormone,GH）基因3＇非翻译区的多态性.结果表明3个群体中3＇非翻译区存在两种长度多态性,分别为320bp和317bp,与GenBank（登录号为：FRU63807）的序列（316bp）有差异;共检测到6种基因型,分别命名为aa、bb、ab、bc、cd、dd,变异频率达4.36%,其中在暗纹东方鲀中检测到四种aa、bb、ab、bc,cd和dd基因型分别只在于星点东方鲀和红鳍东方鲀检测到;7个突变位点中有1处颠换即位点212（T→G）,6处转换即位点120、180、227、265、287（C→T）和位点199（A→G）.     

金钱鱼下丘脑转录组分析及其对雌激素在体注射的响应

为探究XX雌性和XY雄性金钱鱼(Scatophagus argus)下丘脑中的差异表达基因及雌激素(17β-Estradiol,E2)在体注射对XY雄鱼下丘脑中基因表达的影响,开展了转录组测序分析,包括测序数据质控、基因功能注释,差异基因筛选、鉴定和差异基因功能富集分析等,并利用实时定量PCR(Real-time quantitative PCR, qPCR)检测了金钱鱼下丘脑中相关基因的表达。金钱鱼下丘脑转录组共测得Clean reads 275833710,测序质量30(Q30)大于95%, GC含量值大于48%。其中, Ctrl-XX-H vs. Ctrl-XY-H组,共筛选和鉴定了91个差异表达基因,包括36个上调基因和55个下调基因。在Ctrl-XY-H vs. E2-XY-H组,筛选和鉴定了28个差异表达基因,包括11个上调基因和17个下调基因。GO和KEGG富集分析发现,差异表达基因主要显著富集在细胞、单生物过程,膜、膜组分,结合等生物学功能及泛醌和其他萜类-醌生物合成及类固醇激素生物合成通路等。转录组和qPCR结果表明, XX和XY金钱鱼下丘脑中prl、ccna1和hs...     

红鳍东方鲀GPI基因对急性低盐胁迫的响应机制

为了获得红鳍东方鲀Takifugu rubripes葡萄糖-6-磷酸异构酶(Glucose-6-Phosphate Isomerase, GPI)的基因信息及其表达特性, 研究采用RT-PCR和实时荧光定量PCR (qPCR) 技术进行了GPI基因的克隆、组织表达分析及其在急性低盐胁迫下的基因响应研究。结果显示, 所获得的红鳍东方鲀GPI基因序列长1736 bp, 包含一个完整的开放阅读框(Open Reading Frame, ORF)。ORF由1662个核苷酸组成, 编码553个氨基酸; 预测的氨基酸序列中有2个糖异构域(Sugar Isomerase Domains), 不存在信号肽和跨膜结构域。多序列比对结果表明物种间GPI具有较高的保守性。qPCR结果表明: GPI基因mRNA在红鳍东方鲀鳃、肌肉、脑、肠、肝及肾等组织中均有表达, 其中肌肉中表达量最高。在不同盐度胁迫下, 在鳃中, 各低盐组GPI mRNA相对表达量均呈现先升高后降低又回升的趋势; 在肾中, 各低盐组GPI mRNA相对表达量变化趋势各有不同。由此推测, GPI基因在红鳍东方鲀对急性低盐胁迫的响应中发挥一定作用。     

玫烟色棒束孢诱导的小菜蛾免疫响应表达谱分析

【目的】本研究旨在筛选小菜蛾Plutella xylostella应对玫烟色棒束孢Isaria fumosorosea侵染的免疫应答及其网络调控基因,以进一步探讨小菜蛾对玫烟色棒束孢的防御机制。【方法】采用第二代高通量测序技术RNA-seq,对处理后12 h的感染玫烟色棒束孢和健康(平行对照)的小菜蛾4龄幼虫转录组进行了测序,通过生物信息学分析对得到的差异基因进行了功能注释和分类,对差异基因参与的信号通路进行了分析。【结果】在感菌和健康小菜蛾幼虫转录组两个表达谱里,分别获得了12 346 987和12 315 210个clean reads,有60.93%和61.26%的reads分别能比对到参考基因库里,其中完美匹配(perfect match)的比例分别为32.15%和32.73%。共得到351个显著差异表达基因(differentially expressed unigenes,DEUs),上调表达基因有275个,下调表达基因有76个,与免疫防御反应潜在相关的基因有156个。GO(Gene Ontology)富集分析有102个DEUs分布到46个GO term里,KEGG(Kyoto Encyclopedia of Genes and Genomes)pathway富集分析结果显示,有132个DEUs显著富集在13个代谢通路(pathway)里。【结论】这些差异表达基因中,大部分编码潜在的与免疫识别及调控相关的基因,集中在能量代谢、疾病反应和防御反应等相关通路。研究结果为挖掘与玫烟色棒束孢侵染相关的小菜蛾免疫应答候选基因提供了重要数据库,也为阐明小菜蛾对玫烟色棒束孢的免疫机制奠定理论基础。     

小儿脓毒症休克相关基因的筛选及网络构建

为探究脓毒症休克与SIRS的差异表达基因及网络的构建,筛选潜在的核心基因,从GEO数据库下载相关基因表达谱GSE26378,数据分为脓毒症休克与SIRS各29个样本,通过在线软件GCBI对其进行标准化及差异基因筛选;对差异基因进行GO分析;基于KEGG进行功能通路分析以及基因信号网络分析;差异基因共表达网络分析。结果表明:两组中总共有1 456个基因被识别为差异基因(P0.05),与SIRS组相比,脓毒症休克组中有条859条下调基因,597条上调基因。GO功能富集分析显示差异基因主要参与了细胞周期、细胞免疫、细胞代谢。KEGG功能通路分析显示差异基因主要参与了MAPK信号通路、P53信号通路、wnt信号通路、细胞凋亡信号通路,细胞周期受体信号通路等。共表达分析发现基因CCNB1、NUSAP1、OIP5、SHCBP1、ZWINT、TOP2A、DLGAP5等位于网络中央部位,而基因信号网络分析发现基因PLCB1、PIK3CA、STAT3、CAMK2D、PRKCB、CREB1位于网络核心。基因芯片分析有助于发现脓毒症休克与SIRS患儿外周血单核细胞在转录组学上的改变,而生物信息学网络分析有助于发现潜在的靶点。     

红鳍东方鲀干扰素γ基因的原核表达北大核心CSCD

旨在探究一种简单高效的获取重组红鳍东方鲀IFNγ蛋白的方法。提取红鳍东方鲀(Takifugu rubripes)肾脏组织的总RNA,并以其为模板进行RT-PCR扩增干扰素γ(IFNγ)基因序列。将IFNγ成熟肽序列与表达载体p ET-32a(+)相连,成功构建了重组表达载体PET-32a-IFNγ。将其转化入E.coli BL21(DE3)感受态细胞后获得了重组表达IFNγ的基因工程菌。经IPTG诱导,p ET-32a-IFNγ在BL21中得到了表达。通过对表达产物的纯化和Western blotting检测,结果显示红鳍东方鲀IFNγ基因在大肠杆菌中的表达准确,且目的蛋白表达量较高。     

Scurvy in capybaras bred in captivity in Argentine

In order to determine if the absence of vitamin C in the diet of capybaras (Hydrochoerus hydrochaeris) causes scurvy, a group of seven young individuals were fed food pellets without ascorbic acid, while another group of eight individuals received the same food with 1 g of ascorbic acid per animal per day. Animals in the first group developed signs of scurvy-like gingivitis, breaking of the incisors and death of one animal. Clinical signs appeared between 25 and 104 days from the beginning of the trial in all individuals. Growth rates of individuals deprived of vitamin C was considerably less than those observed in the control group. Deficiency of ascorbic acid had a severe effect on reproduction of another population of captive capybaras. We found that the decrease in ascorbic acid content in the diet affected pregnancy, especially during the first stages. The results obtained suggest that it is necessary to supply a suitable quantity of vitamin C in the diet of this species in captivity.     

Changes in lactate dehydrogenase activity in chicken tissues in hyperthyreosis in ontogenesis

The lactate dehydrogenase activity in reactions of lactate oxidation and synthesis was studied in subfractions of the chicken brain, heart and liver at the embryonal, early postembryonal and adult stages of development after thyroxine administration. It has been shown that during embryogenesis thyroxine predominantly enhanced the rate of lactate oxidation in the mitochondrial tissues. A marked increase in the lactate synthesis was found in cytoplasm of the adult chicken tissues. Specificity of enzyme activity alterations was detected in the chicken brain during ontogenesis after thyroxine administration.     

Role for dopamine in malonate-induced damage in vivo in striatum and in vitro in mesencephalic cultures

Defects in mitochondrial energy metabolism have been implicated in the pathology of several neurodegenerative disorders. In addition, the reactive metabolites generated from the metabolism and oxidation of the neurotransmitter dopamine (DA) are thought to contribute to the damage to neurons of the basal ganglia. We have previously demonstrated that infusions of the metabolic inhibitor malonate into the striata of mice or rats produce degeneration of DA nerve terminals. In the present studies, we demonstrate that an intrastriatal infusion of malonate induces a substantial increase in DA efflux in awake, behaving mice as measured by in vivo microdialysis. Furthermore, pretreatment of mice with tetrabenazine (TBZ) or the TBZ analogue Ro 4-1284 (Ro-4), compounds that reversibly inhibit the vesicular storage of DA, attenuates the malonate-induced DA efflux as well as the damage to DA nerve terminals. Consistent with these findings, the damage to both DA and GABA neurons in mesencephalic cultures by malonate exposure was attenuated by pretreatment with TBZ or Ro-4. Treatment with these compounds did not affect the formation of free radicals or the inhibition of oxidative phosphorylation resulting from malonate exposure alone. Our data suggest that DA plays an important role in the neurotoxicity produced by malonate. These findings provide direct evidence that inhibition of succinate dehydrogenase causes an increase in extracellular DA levels and indicate that bioenergetic defects may contribute to the pathogenesis of chronic neurodegenerative diseases through a mechanism involving DA.     

SSTR5 ablation in islet results in alterations in glucose homeostasis in mice

Somatostatin (SST) peptide is a potent inhibitor of insulin secretion and its effect is mediated via somatostatin receptor 5 (SSTR5) in the endocrine pancreas. To investigate the consequences of gene ablation of SSTR5 in the mouse pancreas, we have generated a mouse model in which the SSTR5 gene was specifically knocked down in the pancreatic beta cells (betaSSTR5Kd) using the Cre-lox system. Immunohistochemistry analysis showed that SSTR5 gene expression was absent in beta cells at three months of age. At the time of gene ablation, betaSSTR5Kd mice demonstrated glucose intolerance with lack of insulin response and significantly reduced serum insulin levels. Insulin tolerance test demonstrated a significant increase of insulin clearance in vivo at the same age. In vitro studies demonstrated an absence of response to SST-28 stimulation in the betaSSTR5Kd mouse islet, which was associated with a significantly reduced SST expression level in betaSSTR5Kd mice pancreata. In addition, betaSSTR5Kd mice had significantly reduced serum glucose levels and increased serum insulin levels at 12 months of age. Glucose tolerance test at an older age also indicated a persistently higher insulin level in betaSSTR5Kd mice. Further studies of betaSSTR5Kd mice had revealed elevated serum C-peptide levels at both 3 and 12 months of age, suggesting that these mice are capable of producing and releasing insulin to the periphery. These results support the hypothesis that SSTR5 plays a pivotal role in the regulation of insulin secretion in the mouse pancreas.     

我国葫芦科植物离体培养研究进展

葫芦科植物包括多种瓜类蔬菜,对其进行离体培养研究具有重要的理论和实践意义。综述了国内在葫芦科植物器官培养、体细胞胚胎发生、花药培养、原生质体培养和体细胞杂交及离体遗传转化等方面取得的研究进展,并对葫芦科植物离体培养、遗传转化与育种的前景作了展望。