钱塘江粗唇鱼危细胞色素b基因片段序列分析及系统发生研究

为研究钱塘江粗唇(鮠)与其他鲿科鱼类的亲缘关系,扩增并测定了粗唇(鮠)的细胞色素b基因片段序列.用Blastn分析所获序列与其他鲿科鱼类细胞色素b基因的同源性,用Mega4软件构建系统发生树.     

利用SSR标记分析部分烟草种质的遗传多样性

应用SSR分子标记技术,分析了32份代表性烟草种质的遗传多样性.从38对SSR引物中筛选出8对多态性引物,并对32份烟草材料的基因组DNA进行扩增,共获得93条DNA扩增片断带,其中,多态性带70条,平均多态检出率为75.27%.运用DPS软件进行聚类分析,32个烟草品种被分为4类;其遗传距离变幅在0.31551～1.51978,聚类结果与理论上所期望的结果基本一致,SSR分子标记技术能较好地从分子水平上揭示烟草种质资源遗传背景和亲缘关系.     

Characterization and genomic annotation of polymorphic EST‐SSR loci in Litopenaeus vannamei shrimp

In the present work, EST‐SSR (expressed sequence tag‐simple sequence repeat) loci were obtained by screening 45 000 ESTs from the Pacific white shrimp Litopenaeus vannamei, which was available in a database of the ShEST (Shrimp EST Genome Project) consortium. Fifty‐two of 600 EST‐SSR loci were selected. From this total, 21 EST‐SSRs were polymorphic among 40 individuals and had their gene products ascribed. Two to 20 alleles per locus were detected and the observed heterozygosity ranged from 0.15 to 0.86. Eight loci presented a significant heterozygote deficit after the Bonferroni correction, which was attributed to null alleles. Seven loci were able to have their protein products, molecular functions and biological processes determined. Our results are promising for future studies that relate the levels of these gene polymorphisms with different biological responses to stress in aquaculture.     

Geographical distribution of salmonid alphavirus subtypes in marine farmed Atlantic salmon, Salmo salar L., in Scotland and Ireland

Sequence data from salmonid alphavirus (SAV) strains obtained from farmed marine Atlantic salmon, Salmo salar L. , over a 20-year period between 1991 and 2011 was reviewed to examine the geographical distribution of the genetically defined SAV subtypes in twelve regions across Ireland and Scotland. Of 160 different Atlantic salmon SAV strains examined, 62 belonged to subtype 1, 28 to subtype 2, 34 to subtype 4, 35 to subtype 5 and 1 to subtype 6. SAV subtypes 1, 4 and 6 were found in Ireland, while subtypes 1, 2, 4 and 5 were found in Scotland. In the majority of regions, there was a clear clustering of subtypes, with SAV subtype 1 being the dominant subtype in Ireland overall, as well as in Argyll and Bute in Scotland. SAV subtype 2 predominated in the Shetland and Orkney Islands. The emergence in Atlantic salmon of subtype 2 strains typically associated with sleeping disease in rainbow trout in Argyll and Bute, strongly suggesting transmission of infection between these species, was noted for the first time. SAV subtype 4 was the most common subtype found in the southern Western Isles, while SAV subtype 5 predominated in the northern Western Isles and north-west mainland Scotland. No single strain was dominant on sites in the western Highlands, with a number of sites in this region in particular having more than one subtype detected in different submissions. The significance of these results in relation to aspects of the epidemiology of infection, including transmission, biosecurity and wildlife reservoirs are discussed and knowledge gaps identified.     

猕猴桃钙调蛋白cDNA的克隆及测序

以“秦美”猕猴桃果实呼吸跃变初期ｃＤＮＡ第一链为模板，参考大麦钙调蛋白基因序列合成５′端和３′端引物，利用多聚酶链式反应（ＰＣＲ）合成了完整的猕猴桃钙调蛋白ｃＤＮＡ，克隆并测定了其全序列。结果表明，猕猴桃钙调蛋白基因读码框架内序列由４４４个核苷酸组成，共编码１４８个氨基酸，且与植物领域里发表的大麦及苜蓿的钙调蛋白基因有很高的同源性。在核苷酸序列上与大麦有８６．１６％的同源性，与苜蓿有８５．６９％的同源性，其编码的氨基酸与大麦和苜蓿的不同分别只是一个氨基酸的差异，同源率高达９９．３％．     

星斑川鲽MHCⅡ恒定链Ii基因的克隆和表达特性

为了研究星斑川鲽MHCⅡ类分子的作用及调控机制,实验通过SMART-RACE技术克隆得到了星斑川鲽MHCⅡ恒定链(MHCⅡIi)的全长cDNA序列,其长度为1766 bp,包含135 bp的5′非编码区、837 bp的开放阅读框和794 bp的3′非编码区。该基因共编码279个氨基酸。理论分子量为30.848 ku,等电点为6.89。与已知物种MHCⅡIi进行同源性比对,结果与狼鲈、紫红笛鲷和鳜关系较近,同源性均为79%。利用quantitative realtime PCR(qRT-PCR)技术检测了MHCⅡIi在星斑川鲽不同组织中的表达,以及爱德华氏菌感染前后对该基因在不同组织中表达水平的影响,结果显示:在脾脏、头肾、肝脏、后肠、性腺、心脏、血液、鳃和肌肉组织中,MHCⅡIi mRNA均有表达,但在表达量上有明显差异,脾脏和头肾组织相对表达水平较高,鳃、血液、肌肉、心脏和性腺中的表达水平较低。病原感染后,免疫相关组织脾脏和头肾的表达水平升高最明显,肝脏和后肠的表达水平也略有升高,但变化不明显。本研究可为星斑川鲽MHCⅡ类分子的作用机理提供理论依据,同时为海水养殖鱼类的抗病遗传育种工作提供研究基础。     

微卫星多重PCR分析食蚊鱼系统发育关系

食蚊鱼是全球入侵性物种之一。探讨和研究食蚊鱼快速入侵的原因对进一步的生态学研究有重要意义。实验选取11对多态性微卫星位点,检测了来自欧洲境内3条河流流经的10个不同城市53个家系的755个幼鱼样本。利用微卫星多重PCR技术及Colony统计学软件,分析食蚊鱼系谱关系。食蚊鱼的繁殖能力强,繁殖期的雌性食蚊鱼可一胎产下大量雌雄幼鱼,平均9尾,最多可达72尾;更重要的是实验中发现食蚊鱼存在多重授精现象。结果显示,来自同一父本的幼鱼胚胎最多可达41尾;多重授精加快了食蚊鱼的基因流;系统发育树证实,食蚊鱼来源于遥远的海洋,实验采集的10个群体在系统发育树中主要聚为3支。食蚊鱼入侵性强的关键原因是多重授精,强壮精子具有更强的竞争受精能力,增加了物种的多样性,但受精频率与雌鱼大小可能不成比例。在长距离的迁移中,基因流也在快速变化。     

小熊猫源犬瘟热病毒全基因序列的克隆及序列分析

本研究对军事兽医研究所病毒二室分离驯化后的小熊猫源犬瘟热病毒(canine distemper virus,CDV)基因组进行全序列测定,并对其基因组特征及H基因遗传稳定性进行比较分析。根据GenBank公布的犬瘟热病毒全长基因组序列设计合成17对特异性引物,以小熊猫源犬瘟热病毒总RNA为模板,RT-PCR进行分段扩增,并克隆到pEASY-Blunt Simple载体中,经测序、拼接获得全长cDNA序列。结果显示,小熊猫源犬瘟热病毒全基因序列与GenBank登录号分别为AF014953、AY445077、AY542312、AY466011、AY386316、AF164967、EU716337、EU726268、AY443350、AB474397、GU138403和AY649446的12个不同毒株全基因序列的同源性分别为86.6%、92.4%、92.5%、92.5%、94.3%、96.3%、95.9%、87.1%、94.5%、92.3%、87.4%和94.6%,与标准强毒株A75/17株(AF164967)的亲缘关系最近,全基因同源性达96.3%,但与疫苗株Onderstepoort(AF014953)亲缘关系相对较远,同源性为86.6%。小熊猫源犬瘟热病毒H基因与其他不同地区具有代表性的30株CDV进化树分析显示,小熊猫源犬瘟热病毒属于Asia Ⅰ型,H蛋白中309－311位氨基酸残基所形成的潜在糖基化位点,为疫苗株没有而野毒株所共有的,并且可能与病毒的免疫原性有关。因此,致弱的小熊猫源CDV在预防免疫的针对性上可能强于已有的疫苗株。     

两种大眼蟹线粒体12S rRNA基因序列的比较研究

测定了宽身大眼蟹和日本大眼蟹线粒体12S rRNA基因部分片段的序列,前者为577 bp, 后者为572 bp.二者的核苷酸序列A、T、G、C的含量相似,分别为39.0%、35.7%、16.5%、8.8%;39.3%、 36.2%、 15.8%、8.7%;不包括11处插入/缺失位点,两序列间有75个变异位点,核苷酸差异率为13.18%,其中转换42个、颠换33个, 转换与颠换比约为1.3.对国内外4种大眼蟹长度为331 bp的12S rRNA基因同源序列进行分析,A+T的平均含量为76.6%,明显高于G+C的平均含量,且存有变异位点52个,简约信息位点25个.系统发生树的拓扑结构显示,所有的大眼蟹聚为一支,支持大眼蟹类为一单系.     

中国明对虾肝胰腺细小病毒全基因组的克隆及序列分析

中国明对虾肝胰腺细小病毒是一种单链、线性DNA病毒.本研究利用DNA末端加尾和嵌套PCR首次完成了对虾肝胰腺细小病毒(hepatopancreatic parvovirus,HPV)中国分离株(HPV-Pc)全基因组序列的测定,研究结果表明,HPV-Pc株(GenBank登录号GU371276)的全基因组由6085个核...