Molecular Characterization and Expression Analysis of SKIV Infection of Interferon-Induced Protein with Tetratricopeptide Repeats 1(IFIT1) in Epinephelus lanceolatus

Interferon-induced protein with tetratricopeptide repeats 1(IFIT1), also known as interferon-induced protein 56(IFI56) or Interferon-stimulated protein 56(ISG56), was originally identified as a protein induced upon treatment with interferon and inhibited by viral replication and translational initiation. In this study, Epinephelus lanceolatus IFIT1(ELIFIT1) gene was cloned for the first time. The complete cDNA of El IFIT1 gene includes 2921 nucleotides, and encodes a 437-amino acid(AA) protein. The putative ELIFIT1 protein has 9 TRP domains and is highly similar with IFIT1 proteins in other teleosts. In healthy fish, ELIFIT1 gene was highly expressed in the blood, which indicate its specific function in the peripheral immune system. Its expression was also observed in various immunity-related tissues including spleen, intestine, and kidney, Inducted with spotted knifejaw iridovirus(SKIV), ELIFIT1 gene expression was upregulated in the spleen, kidney, and liver 24 h after induction and reached its peak at 72 h, indicating that ELIFIT1 may play an important role in antivirus. These findings contribute to the understanding of the antiviral regulation of ELIFIT1 gene in teleost.     

Dynamic stress concentration of an elliptical cavity in a semi-elliptical hill under SH-waves

The method of wave-function expansion in elliptical coordinates,elliptical cosine half-range expansion and Mathieu function were applied to obtain an exact analytical solution of the dynamic stress concentration factor(DSCF)around an elliptical cavity in a shallow,semi-elliptical hill.An infinite system of simultaneous linear equations for solving this problem was established by substituting the wave expression obtained by the Mathieu function including the standing wave expression of elliptical lining given herein into the boundary condition obtained by the region-matching method.The finite equations system with unknown coefficients obtained by truncation were solved numerically,and the results in the case of an ellipse degenerating into a circle were compared with previous results to verify the accuracy of the method.The effects of different aspect ratios,incident wave angles and aperture ratios on the dynamic stress concentration around the elliptical cavity were described.Some numerical results,when the elliptical hill was changed into a circular one,were analyzed and compared in detail.In engineering,this model can be regarded as a semi-cylindrical hill with an elliptical cylindrical unlined tunnel under the action of SH waves,and the results are significant in aseismic design.     

南京玄武湖菹草种群的环境效应

为了解玄武湖菹草种群的环境效应,对玄武湖2006年春季水质变化、菹草区内外水质差异进行了监测,并分析了2003-2006年春季水质监测资料,结果表明:菹草区DO高于菹草-水交界区及开阔水域;从菹草区到菹草-水交界区、开阔水域TP、TN逐渐升高,TP均值分别为0.062、0.098、0.105 mg/L,TN均值分别为1.674、2.202、2.412 mg/L;与2003-2005年同期均值相比,2006年3-5月份TP分别下降了15.4%、45%、43.3%,TN分别下降58.2%、43.9%、51.3%,SD保持在65 cm以上;说明菹草在改善湖泊环境和净化水质方面有重要作用.最后提出了改善、维持玄武湖健康稳定水生生态系统的措施和建议.     

基于COI序列的翡翠股贻贝Perna viridis线粒体遗传特性分析及其近缘种间的系统关系探讨

应用通用引物 COIL 1490和 COIH 2198对翡翠股贻贝Perna viridis的性腺和体细胞线粒体DNA进行PCR扩增,获得661bp长度的COI基因片段,经过比对性腺与体细胞的COI片段,发现雄性性腺与体细胞COI基因均为一个单倍型,即体内只有一种线粒体DNA类型,没有发现双单性遗传现象,雌、雄性腺的COI基因片段变异率很低（0.31%）。应用PAUP构建了NJ树、MP树以及贝叶斯法构建了贝叶斯树,对股贻贝属3种间的系统关系进行了分析,结果表明,翡翠股贻贝P. viridis与P. canaliculus 和 P. perna 之间的分化与分歧年代的估算是相吻合的。     

黄斑篮子鱼去毒相关基因的克隆与肝脏组成型表达分析

从基因水平探讨海洋鱼类对海洋藻毒素的去毒分子机理。采用RT-PCR法成功克隆了黄斑篮子鱼Siganus oramin肝脏I时相代谢酶细胞色素P450 1A(CYP1A)、II时相代谢酶alpha型谷胱甘肽S-转移酶(GSTA)和rho型谷胱甘肽S-转移酶(GSTR)、热休克蛋白70 (HSP70)、alpha 1型钠钾ATP酶(ATP1A1)及β-肌动蛋白(beta-actin, ACT)基因cDNA核心序列,序列分别长879 bp、582 bp、588 bp、660 bp、749 bp和554 bp。序列同源性分析发现,属鲈形目的黄斑篮子鱼CYP1A、GSTA和GSTR与同属鲈形目的牙鲆Paralichthys olivaceus、欧洲鲽Pleuronectes platessa、真鲷Pagrus major、鲤形目的斑马鱼Brachydanio rerio 相应氨基酸序列同源性较高,CYP1A和GSTA与非洲爪蟾(两栖类)、鸡(鸟类)、小鼠、大鼠和人(哺乳类)相应氨基酸序列同源性低,这可能与鱼类I、II时相去毒酶基因承担水环境毒素去毒代谢的特殊功能有关;而HSP70、ATP1A和β-肌动蛋白在鱼类、两栖类、鸟类、哺乳类中均有较高的同源性,这可能与这些基因在机体中承担的最基本的生命功能相关。应用半定量RT-PCR的方法,以β-肌动蛋白作为外参照,在指数期增长范围内分别得到了CYP1A、GSTA、GSTR、HSP70和ATP1A1 mRNA与β-肌动蛋白mRNA (%)的比值,确定黄斑篮子鱼肝脏去毒相关基因的组成型表达水平。其中,黄斑篮子鱼肝脏CYP1A、GSTA和GSTR基因组成型表达相对较高,HSP70和ATP1A1基因组成型表达相对较低,这可能与不同基因在黄斑篮子鱼海洋藻毒素去毒分子机理中承担的作用相关,为海洋藻毒素在海洋鱼类中的积聚及代谢去毒分子机制的研究提供了相关数据。     

大菱鲆卵黄原蛋白的分离纯化及Native-PAGE、SDS-PAGE性质鉴定

采用5 mg/kg 17β-雌二醇对大菱鲆(Scophthalmus maximus)进行肌肉注射,一周后尾静脉取血,离心分离获得血浆,经Sephacryl S-300(高分辨)分子筛纯化卵黄原蛋白,对卵黄原蛋白进行常规聚丙烯酰胺凝胶电泳,考马斯亮蓝染色和糖、磷、脂的特征性基团染色.结果表明,17β-雌二醇能诱导大菱鲆产生卵黄原蛋白,以N-ative-PAGE方法计算得到大菱鲆卵黄原蛋白的分子质量为538 ku,以SDS-PAGE方法得出其两种亚基的分子质量分别为100 ku和82 ku.分子筛凝胶过滤能较好地纯化大菱鲆血浆中的卵黄原蛋白.     

栉孔扇贝(♀)×虾夷扇贝(♂)子一代杂种优势的SRAP分析

采用新型的分子标记技术(Sequence-related amplified polymorphism,简称SRAP)对栉孔扇贝(Chlamys farreri)(♀)×虾夷扇贝(Patinopecten yessoensis)(♂)子一代及其双亲的遗传变异进行分析.从不同的引物组合中筛选出9对条带清晰、多态性丰富的组合,9对引物共产生了121条扩增带,其中多态性条带107条,平均每对引物组合产生11.89条多态标记.Shannon信息指数I分别为0.299 9、0.218 9、0.334 3,Nei's基因多样性指数h分别是0.196 4、0.141 1、0.218 9,栉孔扇贝与杂交后代的遗传距离是0.107 4,虾夷扇贝与杂交后代的遗传距离是0.228 7,聚类分析也显示了这一特点.从变异贡献率来看,3个群体的总变异中,有37.31%的变异来源于群体间,62.69%的变异来源于个体间.     

海鞘血细胞培养和细菌污染控制

以海鞘(Ciona savignyi)为研究对象,考察了海鞘血细胞原代培养方法及培养中细菌污染的鉴定和控制.血细胞在培养1 h后贴壁,细胞基本以圆形细胞为主,变形细胞一般呈不规则形.变形细胞存活时间较短,而圆形细胞存活时间较长.对于培养过程中的细菌污染,通过细菌分离、培养和纯培养发现两类菌株检出率较高,均为革兰氏阴性菌.经PCR扩增16S rDNA基因序列片断,结果显示这两类菌株分别属于弧菌属(Vibrio)和施万氏菌属(Shewanella).药敏试验结果显示,弧菌属对氯霉素和环丙沙星等具有较强的敏感性;对施万氏菌属较敏感的药物依次为亚胺培南和氯霉素等.最后比较了几种抗生素组合控制血细胞培养中细菌污染的使用效果,其中氯霉素、亚胺培南加双抗的组合有较好的抑菌效果并对培养细胞的贴壁和生长没有影响.     

海湾扇贝微卫星标记开发及其分离方式分析

利用尼龙膜吸附杂交法构建了海湾扇贝的(AC)_(15)、(AG)_(15)微卫星富集文库,随机挑取3 000个克隆,经菌落原位杂交二次筛选共获得阳性克隆1 268个(42.3%).经序列测定和生物信息学分析后得到521个独立的阳性克隆,其中微卫星506个,小卫星15个.微卫星中,完美型248个,占总数的49.0%;非完美型216个,占42.7%;复合型42个,占8.3%;其中AG/TC重复占70.4%(356个),AC/TG重复29.6%(150个).选择其中的55条序列设计引物,筛选出其中的20对引物检测它们在海湾扇贝CC10家系的双亲和94个子代个体中的分离情况,结果表明:(1) 其中6个位点在母本和子代中发生分离,10个位点在父本和子代中发生分离,4个为双亲共享位点;(2) 2个位点存在无效等位基因;(3) 16个位点符合孟德尔遗传定律,4个位点的子代个体基因型频率偏离孟德尔遗传定律.上述结果表明,这些微卫星引物可以用于构建海湾扇贝的遗传连锁图谱,并且所构建的富集文库适用于微卫星标记的大规模开发.