Nonsynonymous substitution rate (Ka) is a relatively consistent parameter for defining fast-evolving and slow-evolving protein-coding genes

Background  

Mammalian genome sequence data are being acquired in large quantities and at enormous speeds. We now have a tremendous opportunity to better understand which genes are the most variable or conserved, and what their particular functions and evolutionary dynamics are, through comparative genomics.     

水牛CD14 cDNA的克隆、表达及鉴定

目的:克隆水牛白细胞分化抗原14(buffalo cluster of differentiation antigen14,bCD14)基因,表达bCD14蛋白,并进行Western Blot鉴定.方法:采用RT-PCR方法从水牛外周血白细胞中扩增bCD14基因,构建重组质粒pET28a-bCD14,转化入E coli BL21,IPTG诱导表达,对表达蛋白进行可溶性分析及Western blot鉴定.结果:bCD14基因含有一个1 122bp的开放阅读框,编码373个氨基酸;与印度水牛、挪威大鼠和人CD14的cDNA序列同源性分别为97.95%、68.78%、78.60%,氨基酸同源性分别为96.78%、61.27%、72.34%;主要以包涵体形式表达,表达蛋白经Western Blot鉴定,得到了一条约46 kD的特异性条带.结论:该文成功克隆了bCD14基因,表达了bCD14蛋白,为进一步揭示水牛抵抗革兰氏阴性菌感染的免疫机制奠定了基础.     

高体近红鲌的生长与繁殖

高体近红鲌为长江上游的特有鱼类,以2008年5—11月从赤水河赤水市江段采集的540尾高体近红鲌标本为材料,对其生长与繁殖特性进行了研究和分析。结果表明:高体近红鲌鳞片年轮结构呈疏密切割型,年龄特征显著。种群由1—4龄共4个年龄组组成,其中以2龄个体为主。体长分布主要集中在100—160 mm;体重分布主要集中在20.0—50.0 g。总性比为♀∶♂=1.30∶1。体长与鳞径呈直线关系,体长和体重呈幂函数关系且幂指数接近3,基本符合匀速生长类型,体长和体重Von Bertalanffy方程分别为Lt=217.38(1-e-0.2867(t+0.757))和Wt=118.151-e-0.2867(t+0.757)2.8103;生长拐点为2.85龄,拐点对应的体长和体重分别为Lt=140.09 mm,Wt=34.37 g。雌雄初次性成熟年龄均为1龄;繁殖高峰期为6—7月。Ⅳ期雌鱼的绝对繁殖力为950—8655粒,平均值(3087.90±1602.15)粒;体长相对繁殖力FL为10.00—56.20粒/cm,平均值(24.26±10.16)粒/cm;体重相对怀卵量FW为66.08—197.67粒/g,平均值(116.49±32.05)粒/g;卵径频率分布显示高体近红鲌为一次产卵类型。       

浮床生态场空间分布特征

在原位试验的基础上,建立浮床生态场模型.模拟结果表明,浮床生态势值在"场源"处最大,从浮床"场源"向周围水平方向以及向水下垂直方向,随着距离的增加生态势逐渐下降.同时,浮床植物的生物量对生态场效应有显著影响,浮床植物生长越旺盛、生物量越大,生态场效应越强.1 m×1m的浮床,当植物生物量为39.40 kg·m-2时,浮床系统可最远影响到距离浮床1.86 m范围内的水体,并对浮床周围10.90 m2范围内的水体产生作用.该模型可以模拟浮床生态场的空间分布特征及其随植物生长发育的动态变化过程,并可通过模型定量化反映浮床系统在其作用范围内对任意一点的作用强度及其综合影响,该模型的构建将为生态浮床的野外应用提供理论依据.     

产乙醇大肠杆菌的构建及其活性检测方法的改进

adhB和pdc是运动发酵单胞菌产乙醇途径的关键基因,分别编码乙醇脱氢酶和丙酮酸脱羧酶,将添加有聚球藻PCC7942rbcLS基因RBS序列的adhB和pdc基因插入pUC18载体,经双重菌液PCR检验和酶切检验得到分别含有pUC-adhB、pUC-pdc和pUC-adhB-pdc载体的3个重组菌株。活性检测实验表明聚球藻PCC7942的rbcLS基因的RBS序列能有效介导运动发酵单胞菌的adhB和pdc基因在大肠杆菌中表达,摇瓶发酵实验表明重组大肠杆菌的产乙醇能力较出发菌株大幅提升。鉴于乙醛指示平板法存在着对希夫试剂的要求较高、易产生较强的背景色等缺点,对定性检测丙酮酸脱羧酶和乙醇脱氢酶表达菌株的方法做了改进,即:将菌液诱导表达,然后分别添加对应于两种酶的底物,让酶与底物反应0.5至1小时,之后再加希夫试剂进行显色反应,结果表明改进后的方法比乙醛指示平板法更加简便、快速、可靠。     

转病毒来源发夹RNA小麦表现对大麦黄矮病毒的抗性

将大麦黄矮病毒GPV株系的复制酶基因片段和CP基因片段构建成可在植物细胞内表达含有双链复制酶RNA(茎)和反义CP RNA(环)的复合发夹RNA结构, 希望能够诱发植物体针对病毒的RNA干扰作用, 从而达到抗病毒目的。利用基因枪法将该结构导入小麦幼胚愈伤组织细胞后, 通过在幼苗再生阶段进行以叶片为模板的快速PCR来加速阳性植株的筛选过程, 最终共获得基因组整合有外源基因的小麦再生植株21株。对再生植株接种不同剂量的病毒, 其中9株对BYDV-GPV有低度抗性, 表现在低接毒量时无症状, 接毒量提高时发病且严重; 6株具中度抗性, 表现在低接毒量时无症状, 接毒量提高时局部有不严重症状; 6株具高度抗性, 两种情况下均无症状。抗性实验结果表明, hpRNA介导对BYDV的抗性可能受到BYDV含量的影响, 具有剂量效应的特点。     

Condensation of amino acids to form peptides in aqueous solution induced by the oxidation of sulfur(iv): An oxidative model for prebiotic peptide formation

Condensation of amino acids to peptides is an important step during the origin of life. However, up to now, successful explanations for plausible prebiotic peptide formation pathways have been limited. Here we report that the oxidation of sulfur (IV) can induce the condensation reaction of carboxylic acids and amines to form amides, and the condensation reaction of amino acids to form peptides. This might be a general reaction contributing to prebiotic peptide formation.     

Isolation and characterization of <Emphasis Type="Italic">Paenibacillus polymyxa</Emphasis> LY214, a camptothecin-producing endophytic bacterium from <Emphasis Type="Italic">Camptotheca acuminata</Emphasis>

Camptothecin (CPT) is mainly produced and extracted from Camptotheca acuminata and Nothapodytes foetida for pharmaceutical use, i.e., the starting material for chemical conversion to the clinical CPT-type drugs. As the third largest plant anticancer drug, the heavy demand on CPT from global market leads to many research efforts to identify new sources for CPT production. Herein we report the isolation and characterization of a CPT-producing endophytic bacterium Paenibacillus polymyxa LY214 from Camptotheca acuminata. A 10.7 μg l^?1 of CPT was presented in the fermentation broth of P. polymyxa LY214. Its CPT production decreased sharply when the strain of the 2nd generation of P. polymyxa LY214 was cultured and fermented. However, the CPT production remained relatively constant from 2.8 μg l^?1 of the 2nd generation to 0.8 μg l^?1 of the 8th generation of P. polymyxa LY214 under optimized fermentation conditions. A 15- to 30-fold increase of CPT yield was observed when the optimized fermentation conditions, together with the addition of putative biosynthetic precursors of CPT and adsorbent resin XAD16, were applied to ferment the strains of the 7th and 8th generation of P. polymyxa LY214. Bioinformatics analysis of the relative species of P. polymyxa LY214 indicates its potential to produce CPT, which will be helpful to decipher the mysteries of CPT biosynthesis.