水稻幼苗脱黄化过程以及灌浆期茎的蛋白质组学研究

水稻既是我国三大粮食作物之一，又是基因组学研究的模式材料，在生产实践和科学研究中都占有极其重要的地位。基因组学研究取得的巨大成就以前所未有的深度和广度推动了生命科学各个研究领域的飞速发展。水稻基因组的破译是水稻科学研究的重要里程碑，同时也宣告了功能基因组学时代的到来。蛋白质组学是研究细胞内全部蛋白质的动态表达及其相互关系的新兴学科，是功能基因组学研究的重要组成部分和战略制高点。 本论文采用高分辨率的蛋白质双向电泳分离技术和高通量的蛋白质质谱分析技术以及生物信息学等手段，开展水稻灌浆期茎蛋白质组表达模式和水稻幼苗脱黄化过程的比较蛋白质组学研究，探讨茎生长发育规律和水稻应答光信号相关蛋白质及其网络调控机制，是学科前沿与实际应用的有机结合，在科研和生产实践中都具有重要的意义。 首先，分别构建了灌浆期水稻顶端茎段和水稻黄化幼苗的蛋白质组表达谱。并对其中185个目的蛋白点进行了MALDI-TOF/MS分析和数据库检索鉴定。共有149个蛋白质得到了鉴定，蛋白质鉴定的成功率为80.5％。这些被鉴定的蛋白质分属118个基因的表达产物，根据它们功能可以分为13种不同的类别，其中绝大多数为能量产生和代谢以及抗性相关的蛋白质。 在水稻灌浆期顶端茎段表达的蛋白质中，与能量和物质代谢相关的蛋白质例如ATPase、磷酸丙糖异构酶，6－磷酸葡萄糖异构酶等占有很高比例，说明茎段组织中具有很强的代谢活动。与生长发育相关的蛋白质包括beta-tubulins、无机焦磷酸酶（inorganic pyrophosphatase）、液泡质子ATP酶（vacuolar proton-ATPase）以及UDP葡萄糖焦磷酸酶等的大量累积，显示出顶端茎段细胞分裂和生长迅速;同时，贮存多糖和结构多糖也在旺盛合成。G蛋白、GDP释放抑制因子等信号传导蛋白以及苯丙氨酸氨解酶、谷胱苷肽S转移酶（glutathione S-transferase，GST）、抗坏血酸过氧化酶（ascorbate peroxidase，APX）以及超氧化物歧化酶（superoxide dismutase，SOD）等抗性相关蛋白质在该时期丰度表达，表明在灌浆期水稻顶端茎段能够迅速感受并传递外界信号，从而使得其在遭受胁迫时能够立刻启动抗逆防御系统，最大限度地降低不利环境对种子发育的影响。 在黑暗中萌发和生长的水稻黄化幼苗随着光照时间（0~24小时）的延长，能通过双向电泳后检测到的蛋白质逐渐变少，24小时后趋于稳定，相当于正常光照条件下生长的水稻幼苗蛋白质组表达谱。进一步分析表明，在黄化苗中，分解代谢及能量产生相关的蛋白如丙糖磷酸异构酶、琥珀酰辅酶A连接酶、异戊酰辅酶A脱氢酶与ATPase等的表达量比较丰富;另外，还可能启动了脂肪酸的α氧化分解途径，以供黑暗中生长所需的物质和能量。当黄化幼苗光照后，与光合作用及物质合成相关的一些蛋白质表达量增加，而那些分解代谢相关酶类则有所下降。同时，鸟核苷酸结合蛋白β亚基类似蛋白、20S proteasome以及Bowman Birk trypsin inhibitor等信号传递及抗性相关蛋白随着光照时间的延长而减少，说明黑暗胁迫条件下水稻幼苗启动了相关的抗逆途径。叶绿素合成途径中的蛋白酶胆色素原脱氨基酶和金属鳌合酶在脱黄化过程中表达量有所下降，可能是因为叶绿素合成产物具有反馈抑制作用。 本研究首次利用蛋白质组学方法来解析水稻灌浆期茎蛋白质组表达模式和水稻黄化幼苗响应光因子的蛋白质组变化情况，鉴定了一些有价值的蛋白质，并得到了它们的表达特点和相关数据，为更好地理解水稻顶端茎秆的生长特点和功效、水稻应答黑暗胁迫和光形态建成以及光合作用机理等提供了分子证据。

硅基蓝—紫光发光材料及其制备工艺

本发明涉及一种新型硅基蓝一紫光发光材料及其制备工艺，该发光材料是在单晶硅基片层上设有发光层，在发光层上设有薄膜保护层；其制备工艺是先将单晶硅片在高温加气氛下进行热处理；然后将预处理后的单晶硅片注入离子进行掺杂；再用高能重离子对掺杂硅片进行辐照；本发明的性能稳定且发光效率较高，可发出蓝一紫光；其制备工艺代替了传统工艺。

Accelerated evolution of the pituitary adenylate cyclase-activating polypeptide precursor gene during human origin

Pituitary adenylate cyclase-activating polypeptide (PACAP) is a neuropeptide abundantly expressed in the central nervous system and involved in regulating neurogenesis and neuronal signal transduction. The amino acid sequence of PACAP is extremely conserved across vertebrate species, indicating a strong functional constraint during the course of evolution. However, through comparative sequence analysis, we demonstrated that the PACAP precursor gene underwent an accelerated evolution in the human lineage since the divergence from chimpanzees, and the amino acid substitution rate in humans is at least seven times faster than that in other mammal species resulting from strong Darwinian positive selection. Eleven human-specific amino acid changes were identified in the PACAP precursors, which are conserved from murine to African apes. Protein structural analysis suggested that a putative novel Deuropeptide might have originated during human evolution and functioned in the human brain. Our data suggested that the PACAP precursor gene underwent adaptive changes during human origin and may have contributed to the formation of human cognition.

竹叶青蛇毒丝氨酸蛋白酶的分子克隆和序列比较

利用逆转录酶与聚合酶链反应相结合的 RT-PCR 法, 扩增出5个竹叶青(Trimeresurus stejnegeri)蛇毒丝氨酸蛋白酶的 cDNAs; 将扩增的 cDNA 片段克隆入 pGEM-T 载体中, 筛选得到它们的基因, 分别命名为TSSP-1、TSSP-2、TSSP-3、TSSP-4 和 TSSP-5. 经末端终止法测定核苷酸序列, 推导出5个丝氨酸蛋白酶的全序列; 结合纯化的蛋白酶N-末端序列测定结果, 推导 TSSP 2、3和4分别编码凝血酶样酶 stejnobin、纤溶酶 stejnefibrase 1和2. 5个丝氨酸蛋白酶分别含有1～6个N-型糖基结合位点, 表明它们的计算分子量与纯化蛋白表观分子量之间的差异是由糖含量的不同造成, 而其氨基酸序列相似度在60%-90%, TSSP-1 和-2编码的成熟蛋白酶山236个氨基酸残基组成, TSSP-3, -4 和 -5的则山234个氨基酸残基组成, TSSP-1编码的蛋白酶在组成丝氨酸蛋白酶三联体催化活性中心产生了 His~(41)-Arg~(41)的在然突变, 这与其他自然界发现的丝氨酸蛋白酶明显不同。

Study on Mitochondrial DNA Cytochrome b Gene of Chinese sea bass, Lateolabrax sp.

参考鳗鲡等鱼类线粒体 DNA序列进行了中国花鲈线粒体 DNA细胞色素 b基因片断的引物设计、PCR扩增及其序列测定。得到中国花鲈的碱基序列为 4 10 bp,其 A、T、G、C含量分别为 10 1bp(2 4 .6 3% )、112 bp(2 7.32 % )、72 bp(17.56 % )、12 5bp(30 .4 9% ) ,与鳗鲡等其他鱼类相同基因片断序列碱基含量相似。

中国花鲈细胞色素b基因片断的序列研究

参考鳗鲡等鱼类线粒体DNA序列进行了中国花鲈线粒体DNA细胞色素b基因片断的引物设计、PCR扩增及其序列测定。得到中国花鲈的碱基序列为410bp，其A、T、G、C含量分别为101bp（24.63%）、112bp（27.32%）、72bp（17.56%）、125bp（30.49%），与鳗鲡等其他鱼类相同基因片断序列碱基含量相似。

云南保山猪线粒体DNA D loop区序列初步分析

为了解云南保山猪(Baoshan pig)的遗传多样性及其遗传背景，我们测定了19个个体线粒体DNA Dloop高变区1 1 5 363 - 1 5 801片段序列438帅。检测到1。种单倍型，包括8个多态位点，其中5次T/ C转换、1次G/ A转换、1次G/ C颠换和1次A/ T颠换，其A.T.GX碱基的平均含量分别为35.4%.26.9%.13.2%和24.5 %,A+ T含量(62 .3)明显高于G+ C含量(37 .7 %)。对于保山猪的保种及其持续利用有着重要的理论指导意义。

A mitogenomic perspective on the ancient, rapid radiation in the Galliformes with an emphasis on the Phasianidae

Background: The Galliformes is a well-known and widely distributed Order in Aves. The phylogenetic relationships of galliform birds, especially the turkeys, grouse, chickens, quails, and pheasants, have been studied intensively, likely because of their close association with humans. Despite extensive studies, convergent morphological evolution and rapid radiation have resulted in conflicting hypotheses of phylogenetic relationships. Many internal nodes have remained ambiguous. Results: We analyzed the complete mitochondrial (mt) genomes from 34 galliform species, including 14 new mt genomes and 20 published mt genomes, and obtained a single, robust tree. Most of the internal branches were relatively short and the terminal branches long suggesting an ancient, rapid radiation. The Megapodiidae formed the sister group to all other galliforms, followed in sequence by the Cracidae, Odontophoridae and Numididae. The remaining clade included the Phasianidae, Tetraonidae and Meleagrididae. The genus Arborophila was the sister group of the remaining taxa followed by Polyplectron. This was followed by two major clades: ((((Gallus, Bambusicola) Francolinus) (Coturnix, Alectoris)) Pavo) and (((((((Chrysolophus, Phasianus) Lophura) Syrmaticus) Perdix) Pucrasia) (Meleagris, Bonasa)) ((Lophophorus, Tetraophasis) Tragopan))). Conclusions: The traditional hypothesis of monophyletic lineages of pheasants, partridges, peafowls and tragopans was not supported in this study. Mitogenomic analyses recovered robust phylogenetic relationships and suggested that the Galliformes formed a model group for the study of morphological and behavioral evolution.

细胞色素分子疏水性与进化的关系

在先前研究结果的基础上, 通过对细胞色素分子一维结构间疏水相似性的计算, 建立了相应的分子系统树, 并对细胞色素分子间的进化关系进行了探讨。结果表明, 从蛋白质分子的疏水相似性和非线性三维结构来研究分子间的进化关系, 不仅得到了与用其它方法所得到的结果基本一致的结论, 而且还在一定程度上克服了其它一些方法的局限性, 取得了较佳的结果。

Methylation and gene mutation in eukaryotic DNA

5-methylcytosine (m(5)C) as a rare base exists in eukaryotic genomes, which is a normal constitution in many eukaryotic DNA and the existence of m(5)C is a feature of eukaryotic DNA. Under regular physiological conditions, cytosine of eukaryotic DNA is usually methylated. Up to the present, many people consider that the m(5)C may be mutation hotspots by the deamination leading to gene mutation. Our study indicated that the spontaneous mutation caused by the transition of G.C --> A.T, in eukaryotic DNA, may result from the tautomer changing of base pairs and may also be cause by other factor actions, however it could not be caused by the deamination of m(5)C.

HSPC117 deficiency in cloned embryos causes placental abnormality and fetal death

Somatic cell nuclear transfer (SCNT) has been successfully used in many species to produce live cloned offspring, albeit with low efficiency. The low frequency of successful development has usually been ascribed to incomplete or inappropriate reprogramming of the transferred nuclear genome. Elucidating the genetic differences between normal fertilized and cloned embryos is key to understand the low efficiency of SCNT. Here, we show that expression of HSPC117, which encodes a hypothetical protein of unknown function, was absent or very low in cloned mouse blastocysts. To investigate the role of HSPC117 in embryo development, we knocked-down this gene in normal fertilized embryos using RNA interference. We assessed the post-implantation survival of HSPC117 knock-down embryos at 3 stages: E9 (prior to placenta formation); E12 (after the placenta was fully functional) and E19 (post-natal). Our results show that, although siRNA-treated in vivo fertilized/produced (IVP) embryos could develop to the blastocyst stage and implanted without any difference from control embryos, the knock-down embryos showed substantial fetal death, accompanied by placental blood clotting, at E12. Furthermore, comparison of HSPC117 expression in placentas of nuclear transfer (NT), intracytoplasmic sperm injection (ICSI) and IVP embryos confirmed that HSPC117 deficiency correlates well with failures in embryo development: all NT embryos with a fetus, as well as IVP and ICSI embryos, had normal placental HSPC117 expression while those NT embryos showing reduced or no expression of HSPC117 failed to form a fetus. In conclusion, we show that HSPC117 is an important gene for post-implantation development of embryos, and that HSPC117 deficiency leads to fetal abnormalities after implantation, especially following placental formation. We suggest that defects in HSPC117 expression may be an important contributing factor to loss of cloned NT embryos in vivo. (C) 2010 Elsevier Inc. All rights reserved.

Anti-human Immunodeficiency Virus-1 Constituents of the Bark of Poncirus trifoliata

A total of 36 compounds (1-36) were obtained from the stem bark of Poncirus trifoliata including three new prenylated flavonoids, (-)-5,4'-dihydroxy-7,8-[(3 '',4 ''-cis-dihydroxy-3 '',4 ''-dihydro)-2 '',2 ''-dimethylpyrano]-flavone (1), (-)-5,4'-dihydroxy-7,8-[(3 ''-hydroxy-4 ''-one)-2 '',2 ''-dimethylpyrano]-flavone (2), and (-)-5,4'-dihydroxy-7,8-[(cis-3 ''-hydroxy-4 ''-ethoxy-3 '',4 ''-dihydro)-2 '',2 ''-dimethylpyrano]-flavone (3). The new structures were elucidated by means of spectroscopic methods. Compounds 1-20 were evaluated for their anti-human immunodeficiency virus-1 (HIV-1) activity, in which 2 showed significant anti-HIV-1 activity with high therapeutic index (T1) of 143.65.

Chemical Constituents from the Leaves and Stems of Schisandra lancifolia

A new nortriterpenoid, 20-hydroxymicrandilactone D (1) and a novel lignan glycoside, lancilignanside A (2) were isolated from leaves and stems of Schisandra lancifolia, together with three known nortriterpenoids (3-5) and nine known phenolics (6-14). The structures of new compounds 1 and 2 were determined by detailed analysis of their 1D and 2D NMR spectra, and chemical evidences. In addition, compounds 1-2, 6-7, and 9-11 showed anti-human immunodeficiency virus (HIV)-1 activities with 50% effective concentration (EC50) in the range of 3.0-99.0 mu g/ml. Compound 12 was not bioactive in this assay with EC50 more than 200 mu g/ml.

动物模型在HIV-1杀微生物剂有效性和安全性评价中的应用

HIV/AIDS的流行趋势没有减弱的迹象,人们迫切需要新的预防HIV传播的手段。杀微生物剂旨在通过局部用药于阴道或直肠,从而阻止HIV的传播。鉴于目前有大量的杀微生物剂候选物,亟待能够有效评价其有效性及安全性的动物模型。通过比较非灵长类小型动物模型与非人灵长类动物模型在评价HIV杀微生物剂的有效性及安全性上的重要作用,该文总结了评价杀微生物剂有效性及安全性的动物模型的优缺点,同时指出了杀微生物剂研究与发展的方向和建议,希望能够对杀微生物剂的研发有所帮助。