Expression patterns of a circadian clock gene are associated with age-related polyethism in harvester ants, Pogonomyrmex occidentalis

Background  

Recent advances in sociogenomics allow for comparative analyses of molecular mechanisms regulating the development of social behavior. In eusocial insects, one key aspect of their sociality, the division of labor, has received the most attention. Age-related polyethism, a derived form of division of labor in ants and bees where colony tasks are allocated among distinct behavioral phenotypes, has traditionally been assumed to be a product of convergent evolution. Previous work has shown that the circadian clock is associated with the development of behavior and division of labor in honeybee societies. We cloned the ortholog of the clock gene, period, from a harvester ant (Pogonomyrmex occidentalis) and examined circadian rhythms and daily activity patterns in a species that represents an evolutionary origin of eusociality independent of the honeybee.     

Evaluation the effects of dextromethorphan and midazolam on morphine induced tolerance and dependence in mice

The main aim of this study was to evaluate the effects of dextromethorphan and midazolam and their combination on morphine tolerance and dependence in mice. In the present study, different groups of mice were rendered randomly and received morphine (50 mg kg(-1), s.c.), morphine (50 mg kg(-1), s.c.) + Dextromethorphan (25, 50 and 75 mg kg(-1), i.p.), morphine (50 mg kg(-1), s.c.) + midazolam (0.5, 1 and 2 mg kg(-1), i.p.), morphine (50 mg kg(-1), s.c.) + [Dextromethorphan (25 mg kg(-1), i.p. ) + midazolam (0.5 mg kg(-1), i.p.)] once a day for four days. Tolerance was assessed by administration of morphine (9 mg kg(-1), i.p.) on fifth day. Withdrawal symptoms (markers for dependence) was assessed by administration of naloxone (4 mg kg(-1), i.p.) 2 h after co-administration of morphine with either Dextromethorphan or midazolam or their combination. Results showed that pretreatment with Dextromethorphan or midazolam decreased the degree of tolerance and withdrawal symptoms significantly. Additionally co-administration ofDextromethorphan and midazolam couldn't decreased the tolerance and dependence significantly. From these results it may concluded that Dextromethorphan and midazolam alone or in combination could prevent the development of morphine induced tolerance and dependence. These effects can be related to the N-Methyl-D-Aspartate (NMDA) receptor antagonist behavior of Dextromethorphan and GABA-receptor agonist property of midazolam.     

甘蓝型油菜AnnBn1基因的克隆和表达分析

利用电子克隆和RT-PCR技术从甘蓝型油菜抗旱品系Q2中克隆了膜联蛋白(annexin)基因,命名为AnnBn1（GenBank登录号HM244482）,并对其进行了表达分析。AnnBn1的开放阅读框长度为954bp,编码317个氨基酸。序列分析显示,推测AnnBn1基因编码的氨基酸序列含有4个重复的结构域及钙离子结合位点,与拟南芥、番茄、棉花和玉米等物种的膜联蛋白具有较高的同源性。荧光定量PCR结果发现AnnBn1基因在Q2的根、茎、叶、芽等组织中均有表达,而且表达量基本相同。对3叶期幼苗进行10%PEG（聚乙二醇）溶液模拟干旱时发现,干旱胁迫后30h内,AnnBn1基因在茎、芽中的表达量升高了2～4倍,而在叶、根中显著升高了10倍以上。AnnBn1基因表达峰值也具有时空特点,干旱胁迫后20h茎和芽中表达量高,而在30h之后叶和根中表达量高。     

Chronic inhibition of endoplasmic reticulum calcium-release channels and calcium-ATPase lengthens the period of hepatic clock gene Per1

Background

The Magel2 gene is most highly expressed in the suprachiasmatic nucleus of the hypothalamus, where its expression cycles in a circadian pattern comparable to that of clock-controlled genes. Mice lacking the Magel2 gene have hypothalamic dysfunction, including circadian defects that include reduced and fragmented total activity, excessive activity during the subjective day, but they have a normal circadian period. Magel2 is a member of the MAGE family of proteins that have various roles in cellular function, but the specific function of Magel2 is unknown.

Methods

We used a variety of cell-based assays to determine whether Magel2 modifies the properties of core circadian rhythm proteins.

Results

Magel2 represses the activity of the Clock:Bmal1 heterodimer in a Per2-luciferase assay. Magel2 interacts with Bmal1 and with Per2 as measured by co-immunoprecipitation in co-transfected cells, and exhibits a subcellular distribution consistent with these interactions when visualized by immunofluorescence. As well, Magel2 induces the redistribution of the subcellular localization of Clock towards the cytoplasm, in contrast to the nucleus-directed effect of Bmal1 on Clock subcellular localization.

Conclusion

Consistent with the blunted circadian rhythm observed in Magel2-null mice, these data suggest that Magel2 normally promotes negative feedback regulation of the cellular circadian cycle, through interactions with key core circadian rhythm proteins.     

花生线粒体型苹果酸脱氢酶基因AhMMDH1 的结构与表达分析

苹果酸脱氢酶（malate dehydrogenase,MDH）是生物体内十分重要的一类酶,参与植物生长、花粉和果实发育等重要生物过程的调控。当前花生中苹果酸脱氢酶在种子后熟和萌发过程中的作用研究较少。本研究克隆了花生AhMMDH1 基因,其ORF大小为1038bp,编码345个氨基酸,是定位于线粒体的疏水蛋白。三维结构分析发现,AhMMDH1以苹果酸脱氢酶（M链）和乙醛酸前体（G链）的异源二聚体形式存在,其M链在41-47位点出现NAD+结合位点。qRT-PCR分析发现,该基因在花中表达量较高,干种子次之,茎叶中表达痕量;不同发育时期10 ~70 d（新鲜收获种子）种子表达水平均明显低于干种子;种子吸水16h表达量明显升高,胚根突破种皮时（吸水36h）达到最高,随后表达量明显下降。推测该基因在种子后熟和萌发过程中起重要作用。本研究为进一步研究AhMMDH1 的生物功能提供了依据。     

花生DREB类转录因子基因AhDREB3的序列及非生物胁迫下表达分析

DREB类蛋白属于AP2/ERF转录因子家族的一个亚家族,在植物非生物胁迫抗性调控中具有重要功能。为了挖掘花生逆境胁迫相关功能基因AhDREB3,从已公布的花生基因中找到干旱应答元件结合蛋白3（AhDREB3）的编码基因全序列,做编码蛋白的进化分析。根据已知序列设计引物,通过荧光定量PCR检测了该基因在低温、高盐和干旱胁迫下及对外源ABA响应和表达。荧光定量PCR结果显示,AhDREB3基因在花生的叶片和根中对低温没有响应,对高盐（叶片和根）和干旱（根）胁迫有较大响应,说明它可能参与了花生对高盐和干旱胁迫的适应性调控。此外,AhDREB3基因的表达在花生叶片和根中对外源ABA响应变化小,暗示了该基因在花生中可能通过不依赖于ABA的方式起作用。     

以油菜脂肪酸延长酶基因fae1为靶标RNAi载体的构建

通过基因工程手段抑制脂肪酸延长酶基因fae1的表达,以阻止芥酸合成,培育不受高芥酸窜粉影响的低芥酸品种.利用GenBank中的fae1基因序列AF490462为模板设计引物和多聚酶链式反应(PCR)从白菜、甘蓝和甘蓝型油菜的12个不同品种中扩增出长1007bp的fae1基因片段,通过对fae1基因片段DNA序列的测定和序列比较,找到长度为428bp高度保守区作为RNA干涉(RNAi)的靶标区.根据RNA干涉(RNAi)双向表达载体设计原则,将428bp fae1基因片段以正反两个方向插入双向表达载体pMCG161中,两个片段用一个wax基因的内含子连接,植物筛选标记基因采用bar基因,从而构建以油菜fae1基因为靶标的RNAi载体.     

Cloning and expression analysis of HXK1 gene from Jatropha curcas L.

HXK (Hexokinase) gene family and the role of JcHXK1 in Jatropha curcas L. were explored. Totally 4 HXK genes JcHXK1, JcHXK2, JcHXK3 and JcHKL1 were identified and complete ORF of JcHXK1 was cloned. Functional domain, phylogenetic evolution and low-temperature expression characteristics were analyzed. Results showed that full-length JcHXK1 cDNA was 1497 bp, encoding 498 amino acids with molecular weight of 53.81 ​kDa and pI of 5.03. Further phylogenetic evolutionary analysis demonstrated HXK1 protein was clustered into 6 small branches and 2 large branches. Sequence alignment showed that HXK1 contained several conserved glycine residues and hydrophobic channels. Prokaryotic expression vector of JcHXK1 was constructed and 12% SDS-PAGE detection showed that it was highly expressed in E. coli. These research was expected to lay a foundation for further gene functional verification and cold signal transduction mechanism for HXK1 in Jatropha curcas L.     

油菜种子含油量相关的脂肪酶基因BnSDP1克隆与表达分析

通过电子延伸和RACE (cDNA末端快速扩增) 克隆得到一个与油菜种子含油量相关的脂肪酶基因,命名为BnSDP1,其开放阅读框为2 472bp,编码分子量为92kD的蛋白,预测等电点为6.75,具有保守的patatin结构域,跨膜域位于N端和中间,序列分析显示BnSDP1拥有GXSXG脂肪酶的基本特征序列GSSVG。利用RT-PCR技术分析发现,在种子发育的成熟期（即脱水期）,BnSDP1在低油品系种子中的转录水平高于高油品系,在根、茎、叶、花中均有表达,但在根和叶片中表达量高,在根中尤其显著,且受蔗糖诱导上调表达。     

大豆rbcL基因克隆、序列分析及原核表达

利用叶绿体基因组保守性的特征,根据菜豆、豌豆、烟草的rbcL基因序列设计引物,从大豆叶绿体DNA中克隆rbcL基因,全长序列为1488bp,包括1449bp的开放阅读框,编码482个氨基酸。相似性比较显示,此序列与其它10个物种rbcL基因核苷酸的同源性为85.37%～95.31%,氨基酸的同源性为90.87%～96.47%。将该基因与表达载体pET-30a（+）连接,转化大肠杆菌Rosseta感受态细胞,PCR和酶切鉴定筛选阳性克隆,阳性菌液IPTG诱导后经10%SDS-PAGE分析,结果显示：诱导表达出分子量约为60kD的特异融合蛋白。     

矮化大豆GmEXPA5基因的克隆与表达分析

以矮化大豆HK808的顶芽cDNA为模板,根据植物α-膨胀素基因保守区设计简并引物,并克隆保守区片段,结合RACE 技术克隆得到一个新的膨胀素全长基因,将其命名为GmEXPA5（登录号为JN207916.1）,该基因全长1 233bp,其中开放阅读框636bp,编码211个氨基酸;推导的氨基酸序列经分析发现,该序列含有两个膨胀素保守域domain1（expansin EG45）和domain2（expansin CBD）,无明显的信号肽序列,属于α膨胀素亚家族。构建pEASY-Blunt E1-GmEXPA5重组质粒,获得稳定的原核表达体系,IPTG 诱导后SDS-PAGE 结果表明所表达蛋白与预期蛋白大小一致。实时荧光定量PCR（real-time PCR）表达分析表明GmEXPA5基因在矮化大豆HK808与野生型大豆东农42两种材料的不同器官中均有表达,而在野生型茎尖及茎部的表达量显著高于矮化大豆,推测该基因与大豆矮化性状的形成可能有一定的关系。     

大豆膨胀素GmEXPA4基因的克隆及表达分析

根据其他植物膨胀素基因保守区设计简并引物,以大豆矮化突变体HK808幼芽的cDNA为模板,克隆到530bp的膨胀素基因保守区片段,并结合RACE 技术克隆得到expansin基因全长序列,将其命名为GmEXPA4基因。生物信息学分析结果表明,该基因属于膨胀素中的α-expansin亚家族。GmEXPA4基因的实时荧光定量相对表达分析表明,该基因在东农42及其矮化突变体HK808中的各个时期都有表达,且与植物的生长量密切相关。     

Comparative effects of scopoletin and cyanide on rat brain, 1: Histopathology

Four week old male Wistar rats were used to study the effects of scopoletin and cyanide on the histopathology of rat brain. The rats were divided into a control and three experimental groups (2–4) and fed rations containing 0.07 g scopoletin/100 g, 0.07g scopoletin + 1.8 mg cyanide/100 g and 1.8 mg cyanide/100 g, respectively. These levels of scopoletin and cyanide corresponded to levels found in a processed cassava diet. The first group was fed the same ration as the others but without scopoletin and cyanide. The rats were fed these rations for twelve months. Rats from each group were sacrificed at the third, sixth, ninth and twelfth months; the relative brain weight of the rats (% of body weight) and histology of their brains were studied. The lipid peroxide levels of the rat brains were also studied at the twelfth month. The results showed that the relative brain weights of the rats fed scopoletin + cyanide were significantly (p<0.05) less than that of the control from the third month. There were no significant changes in the lipid peroxide levels of the rat brains in the various groups. Histological examination of the brains of the rats sugested that scopoletin is involved in the pathogenesis of the neuropathy seen in cassava consuming populations.     

甘蓝型油菜BnNAC61基因的克隆及表达特征分析

NAC(即NAM、ATAF和CUC)转录因子参与植物生长、发育、衰老和多种逆境胁迫反应的调控.为阐释甘蓝型油菜BnNAC61的表达特征,本研究采用RT-PCR方法,从甘蓝型油菜总cDNA中克隆BnNAC61基因.生物信息学分析表明,其CDS全长846 bp,编码281个氨基酸,N端含有NAM保守结构域;其启动子区存在W...