Horizontal Transfer of Archaeal Genes into the Deinococcaceae: Detection by Molecular and Computer-Based Approaches

Members of the Deinococcaceae (e.g., Thermus, Meiothermus, Deinococcus) contain A/V-ATPases typically found in Archaea or Eukaryotes which were probably acquired by horizontal gene transfer. Two methods were used to quantify the extent to which archaeal or eukaryotic genes have been acquired by this lineage. Screening of a Meiothermus ruber library with probes made against Thermoplasma acidophilum DNA yielded a number of clones which hybridized more strongly than background. One of these contained the prolyl tRNA synthetase (RS) gene. Phylogenetic analysis shows the M. ruber and D. radiodurans prolyl RS to be more closely related to archaeal and eukaryal forms of this gene than to the typical bacterial type. Using a bioinformatics approach, putative open reading frames (ORFs) from the prerelease version of the D. radiodurans genome were screened for genes more closely related to archaeal or eukaryotic genes. Putative ORFs were searched against representative genomes from each of the three domains using automated BLAST. ORFs showing the highest matches against archaeal and eukaryotic genes were collected and ranked. Among the top-ranked hits were the A/V-ATPase catalytic and noncatalytic subunits and the prolyl RS genes. Using phylogenetic methods, ORFs were analyzed and trees assessed for evidence of horizontal gene transfer. Of the 45 genes examined, 20 showed topologies in which D. radiodurans homologues clearly group with eukaryotic or archaeal homologues, and 17 additional trees were found to show probable evidence of horizontal gene transfer. Compared to the total number of ORFs in the genome, those that can be identified as having been acquired from Archaea or Eukaryotes are relatively few (approximately 1%), suggesting that interdomain transfer is rare.     

Chickpea facilitates phosphorus uptake by intercropped wheat from an organic phosphorus source

Pot experiments were conducted to investigate interspecific complementation in utilization of phytate and FePO₄ by plants in the wheat (Triticum aestivum L.)/chickpea (Cicer arietinum L.) intercropping under sterile and non-sterile conditions. The pots were separated into two compartments by either a solid root barrier to eliminate root contact and solute movement, by a nylon mesh (30 M) to prevent root contact but permit solute exchange, or not separated between the compartments. Wheat plants were grown in one compartment and chickpea in the other. Two P sources were tested at 60 mg P kg^–1 soil (sodium phytate or FePO₄). Under non-sterile conditions, the biomass of wheat was significantly greater when the roots were intermingled with chickpea than when the roots were separated from chickpea roots by a solid root barrier or nylon mesh. When phytate–P was applied, P concentrations in wheat (2.9 g kg^–1 in shoots and 1.4 g kg^–1 in roots) without root barrier between the two species were higher than those in the treatments with nylon mesh or with the solid root barrier separation (1.9 g kg^–1 in shoots and 1.0 g kg^–1 in roots). In contrast, P concentrations in wheat supplied with FePO₄ were similar between the root separation treatments. There was no significant difference in P uptake by chickpea between the P sources or between the root separation treatments, except that P uptake was greater in the phytate treatment with the root barrier. Total P uptake from phytate was increased by 25% without root separation compared to the root separation treatments. Under sterile conditions and supply of phytate–P, the biomass of wheat was doubled when the roots were intermingled with chickpea and increased by a third with the nylon mesh separation compared to that with the solid root barrier. Biomass production in wheat at various treatments correlated with P concentration in shoot. Biomass production and P concentration in chickpea were unaffected by root separation. Total P uptake by plants was 68% greater with root intermingling and 37% greater with nylon mesh separation than that with the solid root barrier. The results suggest that chickpea roots facilitate P utilization from the organic P by wheat.     

利用全基因组连锁不平衡估计中国荷斯坦牛有效群体大小

有效群体大小是群体遗传学研究的一个重要内容,有助于我们更清楚地了解群体的遗传变异、进化和复杂性状的遗传机制等。随着高密度SNP标记的出现,越来越多的研究利用SNP标记间连锁不平衡估计有效群体大小。文章采集北京地区中国荷斯坦牛2 093个样本,并利用牛SNP芯片(Illumina BovineSNP50,含5 4001 SNPs)进行基因型测定,估计不同世代中国荷斯坦牛的有效群体大小。质量控制标准设定为SNP检出率0.95,最小等位基因频率>0.05,样本检出率0.95,哈代温伯格平衡检验显著性水平P<0.0001。经过质量控制,共1 968个样本和38 796个SNPs用于连锁不平衡分析。文章选取SNP间距0.1、0.2、0.5、1、2、5、10、15(Mb),估计中国荷斯坦牛在4世代之前有效群体大小。结果表明,中国荷斯坦牛的有效群体呈逐代下降趋势,至4世代前,中国荷斯坦牛平均有效群体为45头左右。     

Localization and synthesis of an antigenic determinant of herpes simplex virus glycoprotein D that stimulates the production of neutralizing antibody.

An antigenic determinant capable of inducing type-common herpes simplex virus (HSV)-neutralizing antibodies has been located on glycoprotein D (gD) of HSV type 1 (HSV-1). A peptide of 16 amino acids corresponding to residues 8 to 23 of the mature glycoprotein (residues 33 to 48 of the predicted gD-1 sequence) was synthesized. This peptide reacted with an anti-gD monoclonal antibody (group VII) previously shown to neutralize the infectivity of HSV-1 and HSV-2. The peptide was also recognized by polyclonal antibodies prepared against purified gD-1 but was less reactive with anti-gD-2 sera. Sera from animals immunized with the synthetic peptide reacted with native gD and neutralized both HSV-1 and HSV-2.     

动态检测血管紧张素Ⅱ刺激后STAT3信号核转位及其与衰老的关系

为了动态观察复制性衰老细胞中血管紧张素Ⅱ(AngⅡ)激活人信号转导与转录活化因子3(hSTAT3)信号转导途径的核转位情况及该途径在细胞衰老过程中的变化。将载体pMS1-hSTAT3上的目的基因STAT3序列亚克隆到pEGFP-C3报告载体中,构建出pEGFP-hSTAT3质粒选择人胚肺二倍体成纤维细胞WI-38细胞株进行细胞培养,通过脂质体(Effectene)转染的方法,将pEGFP-STAT3质粒分别转染至19代,42代WI-38细胞中;在激光共聚焦显微镜下动态观察血管紧张素Ⅱ激活STAT3的核转位的变化及不同代数的差别．结果显示AngⅡ刺激细胞后,出现活化的STAT3在胞核内聚集现象,并且在19代细胞中15分钟开始出现,30—45分钟左右时达到高峰;42代细胞中30分钟左右开始出现,50—60分钟左右达到高峰。因此我们认为AngⅡ刺激WI-38细胞,能出现hSTAT3信号转导的核内集聚现象;并且随着WI-38细胞传代数的增加,STAT3信号转导入核时间延迟。说明随着细胞的复制性衰老,通过STAT3通路转导的信号活性逐渐降低,最终影响细胞的增殖,可能是促进细胞衰老的原因之一。     

植物光诱导延迟荧光测量的影响因素研究

光合作用是地球上最重要的化学反应,植物内源性光诱导延迟荧光是光合作用原初过程中光系统Ⅱ作用中心P680处电荷分离效率的内在探针。延迟荧光除了受植物本身及其生长发育状况有关外,还受到其他很多环境及测量方面的影响,所以为了更好地利用延迟荧光特性技术研究植物生理特性,就必须对测量参数指标做合理的优化。本文从影响延迟荧光的激发光源的光强,激发时间及外界环境温度出发,研究延迟荧光的变化特性,为延迟荧光在植物生理特性方面的研究提供合理的理论依据。     

Molecular genetic characterization of rice seed lipoxygenase 3 and assessment of its effects on seed longevity

Lipoxygenases (LOXs) are enzymes involved in lipid peroxidation. Here we reported the identification, molecular and functional characterization of the gene encoding rice (Oryza sativa L.) seed LOX3 (sLOX3). Via a map-based cloning strategy we identified Os03g0700400 as the candidate gene encoding sLOX3. Further functional complementary test and biochemical characterization of the recombinant Os03g0700400 protein verified the identification. The sLOX3 gene was highly expressed in roots, moderately in embryos and very weakly in leaves, leaf sheaths and stems. Transient expression experiment (in rice protoplasts) and subsequent laser confocal microscopic analysis demonstrated that the sLOX3 protein was localized into the cytosol. We next showed that overexpression of sLOX3 in a japonica sLOX3-normal rice cultivar, Wuyunjing 7 accelerated the decrease of seed germination ability when the seeds were routinely stored, which demonstrated that sLOX3 had a negative effect on seed longevity (storability). Meanwhile, an increased occurrence of embryo decay was observed in the same transgenic seeds, suggesting that sLOX3 might negatively affect seed longevity by facilitating colonization of particular seed pathogens. Our result forwarded the understanding of the effects of 9-LOX on rice seed longevity.     

花生根瘤菌类菌体氢氧化体系的研究

花生根瘤菌类菌体含有吸H₂酶,以O₂或亚甲蓝为电子受体均表现吸H₂活性。类菌体还原减氧化的吸收差示光谱在424、500、520、550、560、585及595 nm处呈现吸收峰,表明细胞色素c、b、和o参与H₂的氧化过程。CN-明显影响520、550、560 nm处的吸收峰,意味着有些细胞色素c和b对CN^-敏感,在H₂代谢中充当末端氧化酶的作用。抗霉素A、HONO、CN^-、N₃^-和DBMIB均强烈抑制吸H₂活性,但鱼藤酮不抑制吸H₂活性。说明细胞色素b、c、a和泛配参与H₂氧化的电子传递,而NADH脱氢酶不参与。花生根瘤菌类菌体的H₂氧化系统是个复杂具分支的电子传递体系。     

单克隆人胰腺干细胞分化特性的研究

研究1例来源于4月龄男性流产胎儿胰腺组织的单克隆人胰腺干细胞（monoclonal human pancreatic stem cell,mhPSC）系的体内外分化特性。将mhPSCs接种在铺有0．1％明胶的培养皿内,扩增培养3d后,加高糖DMEM诱导液诱导培养25d。相差显微镜下．观察细胞生长状况。采用双硫腙染色法、RT—PCR及葡萄糖刺激释放胰岛素和C肽实验．对体外定向诱导mhPSCs分化为功能性胰岛进行检测。将mhPSCs悬液注射在成年雄性裸鼠腹股沟皮下．注射30d时,取出移植物,采用SP法进行免疫组织化学反应,以检测mhPSCs的体内自然分化潜能。体外扩增培养,mhPSCs贴壁生长,呈多角形上皮样。生长至单层．呈“铺路石”状。体外定向诱导,细胞逐渐由多角形变成圆形,并聚集成类胰岛。诱导培养15d时．形成的类胰岛中少数细胞分化为B细胞,双硫腙染色阳性。诱导培养25d时,多数细胞分化为8细胞,双硫腙染色阳性,转录表达胰岛素的mRNA。用不同浓度葡萄糖刺激．诱导胰岛不仅释放胰岛素和C肽,而且其释放量随糖刺激浓度升高显著增加（0．01〈P〈0．05）。体内分化实验显示,mhPSCs在裸鼠背部形成类畸胎瘤。类畸胎瘤易与裸鼠分离,色白,血管丰富。显著表达pdx1、胰岛素、胰高血糖素、CK、MBP及NF蛋白。该研究结果证实单克隆人胰腺干细胞系体外定向诱导分化为包含大量β细胞的功能性类胰岛,在体内自然分化为胰岛、上皮及神经组织细胞。