Adaptive evolution of rbcL in Reaumuria soongarica (Tamaricaceae)

In the field of phylogenetic analyses, the rbcL gene encoded large subunit Ribulose-1,5-biphosphate carboxylase/oxygenase (Rubisco, EC4.1.1.39), which plays a crucial role in the process of photosynthesis for most terrestrial plants, has been considered to be conserved; however, recent controversy regarding rbcL conservation has appeared since it was proposed to be under natural selection within all principal lineages of land plants. In this study, by examining the variation of DNA and protein sequences among 17 species in the family Tamaricaceae, three nonsynonymous mutations were identified to be under positive selection. The favored sites were located in the alph-helix domains of Rubisco, with decreased hydrophobicity and increased entropy, which could facilitate CO₂ penetration into the active site of Rubisco. We also found that the expression level of rbcL in different genotypes of Reaumuria soongarica shifted in response to various stresses such as drought, temperature, salt, and light. This study not only sheds light on the functional/structural features of Rubisco in the evolution scenarios from C₃-like into C₄ in Tamaricaceae but also provides useful information on directing genetic performance to enhance photosynthesis efficiency of desert plants for sustaining fragile desert ecosystems; furthermore, it promotes the ability to cope with desert aridification and global warming.     

Genomic cloning and expression of vitellogenin gene from the self-fertilizing fish Rivulus marmoratus (Cyprinodontiformes, Rivulidae)

We cloned the vitellogenin gene from the self-fertilizing fish Rivulus marmoratus, and sequenced 12,326 bp. The number of exons of R. marmoratus and rainbow trout vitellogenin genes were different, and also the splicing junctions are different throughout most of the exons and introns but the amino acid similarity of R. marmoratus vitellogenin gene to other species was rather high. In promoter region of R. marmoratus vitellogenin gene, there were several E2 binding sites and the estrogen response element (ERE). We discuss here the gene structure and expression of R. marmoratus vitellogenin gene.     

鰶亚科两种鱼类的线粒体16S rRNA基因片段序列的比较研究

对鰶亚科两种鱼类的线粒体16S rRNA基因片段进行了PCR扩增和序列测定,分析比较了两种间的序列差异。斑鰶(Konosirus punctatus)16S rRNA基因片段长度为572bp,在3个个体中检测到2个单倍型,花鰶(Clupanodon thrissa)序列长度为575bp,两种间存在30个核苷酸位点(5.2%)的变异。基于木村双参数进化模型得到两种间的遗传距离为0.043。以太平洋鲱和寿南小沙丁为外群构建的NJ树表明,斑鰶和花鰶的亲缘关系较近,而与美洲真鰶(Dorosoma cepedianum)的亲缘关系较远。     

海藻乙醇提取物抗菌活性的研究

分属绿藻门、红藻门、褐藻门的海藻样品于 2 0 0 0年 5月采于山东青岛和威海附近沿海 ,将海藻样品的乙醇提取物进行薄层层析分离和抗大肠杆菌、金黄色葡萄球菌活性测定。结果表明 ,抗大肠杆菌活性较强的海藻样品有 :鸭毛藻、酸藻、松节藻和孔石莼的水相 ;抗金黄色葡萄球菌活性较强的提取物有 :鸭毛藻、海黍子、松节藻、小粘膜藻的乙酸乙酯相以及酸藻的水相。从不同门类海藻的抗菌活性看 ,褐藻最强 ,红藻次之 ,而绿藻活性最弱。薄层层析分离得到 1 6个层析带具有抗金黄色葡萄球菌活性 ,1 1个层析带具有的抗大肠杆菌性。其中褐藻门的酸藻、海黍子的 3个层析带具有很强的抗金黄色葡萄球菌活性 ,酸藻的 2个层析带也能强烈地抑制大肠杆菌的生长。在同样的实验条件下 ,对常见抗生素进行的对照实验表明 ,这 5条层析带的活性比常见的抗生素高。     

海洋经济贝类育种研究进展

本文简要回顾了近些年来海洋经济贝类育种现状 ,综述了传统的杂交育种和各种现代生物技术如多倍体、转基因和分子标记技术在贝类育种中所取得的成就 ,特别是近年来分子标记技术应用于贝类杂种优势预测 ,群体遗传结构分析和标记辅助选择等。为新品种的培育提供了高效而可靠的手段。该技术的应用可望在较短的时间内改变目前海洋贝类品种培育的落后局面     

最小弧菌热不稳定型溶血素表达载体的构建和高效表达

根据已发表的最小弧菌热不稳定型溶血素（Vibrio mimicusheat-labile hemolysinVmhA）核苷酸序列,设计和合成一对特异性引物,以最小弧菌96-6-3的基因组DNA为模板,PCR扩增得到热不稳定型溶血素基因片段,克隆于表达载体PQE-2/VmhA中,转化大肠杆菌M15,经IPTG诱导,表达出预期大小50.2KD的融合蛋白.Westernblot检测显示,该蛋白与抗最小弧菌热不稳定型溶血素兔血清呈阳性反应,表明该蛋白具有溶血素的免疫原性,可作为基因工程疫苗的候选抗原.     

聚合酶链反应(PCR)检测花鲈鳗弧菌感染

从海洋鱼类重要病原菌———鳗弧菌基因库中选择金属蛋白酶基因为目标检测基因 ,以此设计一对引物 ,建立聚合酶链反应 (PCR)检测鳗弧菌的方法 ,并对此方法的灵敏性和特异性进行了研究 ,结果表明该方法对鳗弧菌的检测快速、灵敏。对人工感染鳗弧菌的花鲈组织样品进行检测 ,该方法能识别组织样品中极微量的鳗弧菌。     

三疣梭子蟹线粒体DNA 16S rRNA和COI基因片段序列的比较研究

本文采用 PCR扩增和序列测定等技术 ,对三疣梭子蟹 (Portunustrituberculatus)线粒体DNA1 6Sr RNA和 COI基因片段进行了初步研究。经 PCR扩增和序列测定 ,分别得到 1 6Sr RNA和 COI2个基因片段的碱基序列 ,其中 1 6Sr RNA基因片段的大小为 566bp,碱基 A,T,G,C的含量分别为 35.1 6% ,34.45% ,1 2 .37%和 1 8.0 2 % ;COI基因片段的大小为 658bp,碱基 A,T,G,C的含量分别为 36.63% ,2 6.44% ,2 0 .52 %和 1 6.41 %。在 2种基因片段中 ,AT含量均明显高于 GC含量 ,这与果蝇、虾类、蟹类等无脊椎动物的 1 6Sr RNA和 COI基因片段研究结果相似。通过对三疣梭子蟹 1 6S r RNA和 COI2个基因片段遗传特征的研究 ,发现其种内变异较低 ,在 3个样本中 1 6Sr RNA基因片段序列完全一样 ,COI基因片段中也只有 2个 T/ C位点转换。另外 ,比较本研究所得序列与 Gen Bank中梭子蟹科 5个属的 1 6Sr RNA基因片段序列后 ,发现其聚类结果与传统分类相一致。     

条斑紫菜丝状孢子体cDNA文库构建及抗病相关基因鉴定

经过微量总 RNA抽提、c DNA合成、c DNA扩增和克隆等步骤 ,构建了微量条斑紫菜丝状孢子体 c DNA文库 ,并进行了小规模表达序列标签分析。从 1 70条标签序列中鉴定出 6条抗病相关基因序列。这些序列可用于研制抗病单核苷酸多态性标记 ,辅助紫菜抗性遗传改良。