17α-甲基睾酮对稀有鮈鲫精巢DNA甲基转移酶基因表达的影响

[目的] 以稀有鮈鲫雄鱼为研究对象,探讨17α-甲基睾酮（17α-methyltestosterone,MT）对稀有鮈鲫精巢甲基转移酶（DNMT）基因相对表达量的影响。[方法] 采用不同浓度MT（25、50和100 ng/L）处理稀有鮈鲫雄鱼7、14和21 d,处理结束时,每组随机解剖10条鱼,取精巢组织,Trizol一步法提取总RNA,实时荧光定量PCR（qRT-PCR）检测DNA甲基转移酶基因（dnmts）相对表达量。[结果] MT处理稀有鮈鲫雄鱼7 d,25 ng/L MT处理组（P<0.05）、50 ng/L MT处理组（P<0.01）和100 ng/L MT处理组（P<0.05）精巢dnmt3基因相对表达量显著（或极显著）降低;25 ng/L MT处理组和50 ng/L MT处理组精巢dnmt5基因相对表达量显著（P<0.05）降低;50 ng/L MT处理组精巢dnmt7基因相对表达量显著（P<0.05）降低。MT处理稀有鮈鲫雄鱼14 d,100 ng/L的MT处理组dnmt3、dnmt5、dnmt6、dnmt7基因相对表达量显著（P<0.05）升高;50 ng/L的MT处理组dnmt1基因相对表达量极显著（P<0.01）升高。MT处理稀有鮈鲫雄鱼21 d,50 ng/L MT处理组和100 ng/L MT处理组dnmt1基因相对表达量显著（P<0.05）升高;100 ng/L MT处理组dnmt8基因相对表达量极显著（P<0.01）升高。[结论] MT可以干扰稀有鮈鲫精巢DNA甲基转移酶基因dmmts的mRNA表达。MT对DNMTs酶活性的影响以及是否可以通过改变DNA甲基化水平对稀有鮈鲫精子的发生产生影响有待研究。     

猬迭宫绦虫线粒体cox3基因的克隆及序列分析

本研究旨在阐明猬迭宫绦虫湖南分离株线粒体细胞色素c氧化酶第Ⅲ亚基（cox3）基因部分序列（pcox3）的遗传变异情况。利用聚合酶链反应（PCR）扩增猬迭宫绦虫的pcox3,应用ClustalX 1.81程序对序列进行比对,同时利用DNAStar 5.0中的Megalign程序进行同源性分析,再用Phylip 3.67程序MP法绘制系统发育树,并与GenBank中已知猬迭宫绦虫相应基因序列进行比对分析。结果显示,所获得的pcox3序列长度一致,均为264 bp,湖南分离株与已知猬迭宫绦虫位于同一分支,来自湖南省的猬迭宫绦虫pcox3序列有微小差异。本研究结果为猬迭宫绦虫进一步的分类、鉴定和遗传变异研究奠定了基础     

应用微卫星DNA标记对BWEL-SPF鸡种群的群体遗传学分析

本研究利用14个鸡微卫星DNA标记位点,采用DNA测序仪和荧光素标记分子内标,建立了鸡分子遗传学的检测方法,对F15世代BWEL-SPF鸡种群8个家系群体内和群体间的遗传结构进行了分析。结果表明,被检位点35.7%(5/14)的基因型趋于纯合,家系内等位基因的变异程度显著降低。F-统计量检验结果表明,家系间的遗传分化均达到了极显著水平(P0.001),群体间的变异占到了总遗传变异的12.9%。     

DNA barcoding for identification of cryptic species in the field and existing museum collections: a case study of Aethomys and Micaelamys (Rodentia: Muridae)

DNA barcoding has been proposed as a method for species identification. However, this method has been criticised for its over-reliance on a single mitochondrial gene. In this study, four mitochondrial gene regions and one nuclear gene region were used to investigate their different abilities to identify tissue associated with museum specimens of Aethomys chrysophilus, Aethomys ineptus and Micaelamys namaquensis. Aethomys chrysophilus and the more recently elevated A. ineptus are indistinguishable on morphological grounds; however, their ranges are largely parapatric with only one syntopic locality currently known. All of the mitochondrial gene regions were able to separate M. namaquensis from A. chrysophilus and A. ineptus, but they varied in their abilities to resolve differences between A. chrysophilus and A. ineptus. The sequence results identified a specimen from KwaZulu-Natal that was misclassified and should have been identified as A. ineptus. Seven specimens that had not been reclassified following the elevation of A. ineptus to species level were identified as A. ineptus. Individuals of A. chrysophilus from Malawi could not be classified as either A. chrysophilus or A. ineptus, and may be a hybrid or a new, distinct species. This study indicates that DNA barcoding may be used to separate M. namaquensis from A. chrysophilus and A. ineptus, and although it was not able to separate A. chrysophilus and A. ineptus, it did indicate specimens from Malawi may be a new cryptic species.     

First Steps Towards an Understanding of a Mode ofCarcinogenic Action for Vanadium Pentoxide

Inhalation of vanadium pentoxide clearly increases the incidence of alveolar/bronchiolar neoplasms in male and female B6C3F1 mice at all concentrations tested (1, 2 or 4 mg/m³), whereas responses in F344/N rats was, at most, ambiguous. While vanadium pentoxide is mutagenic in vitro and possibly in vivo in mice, this does not explain the species or site specificity of the neoplastic response. A nose-only inhalation study was conducted in female B6C3F1 mice (0, 0.25, 1 and 4 mg/m³, 6 h/day for 16 days) to explore histopathological, biochemical (α-tocopherol, glutathione and F2-isoprostane) and genetic (comet assays and 9 specific DNA-oxo-adducts) changes in the lungs. No treatment related histopathology was observed at 0.25 mg/m³. At 1 and 4 mg/m³, exposure-dependent increases were observed in lung weight, alveolar histiocytosis, sub-acute alveolitis and/or granulocytic infiltration and a generally time-dependent increased cell proliferation rate of histiocytes. Glutathione was slightly increased, whereas there were no consistent changes in α-tocopherol or 8-isoprostane F2α. There was no evidence for DNA strand breakage in lung or BAL cells, but there was an increase in 8-oxodGuo DNA lesions that could have been due to vanadium pentoxide induction of the lesions or inhibition of repair of spontaneous lesions. Thus, earlier reports of histopathological changes in the lungs after inhalation of vanadium pentoxide were confirmed, but no evidence has yet emerged for a genotoxic mode of action. Evidence is weak for oxidative stress playing any role in lung carcinogenesis at the lowest effective concentrations of vanadium pentoxide.     

鸡柔嫩艾美耳球虫3-1E和CDPK双抗原DNA疫苗的免疫保护效果观察

将构建的含鸡柔嫩艾美耳球虫(Eimeria tenella)CDPK和3-1E双抗原的真核重组质粒proEC,在14日龄和21日龄分别免疫AA肉鸡,28日龄感染5×104个E.tenella孢子化卵囊,观察其免疫保护效果.结果显示,攻虫后1周,与proC组相比,proEC组能显著增强淋巴细胞转化和血清抗体水平(P＜0.05);与proE组和proC组相比,proEC组卵囊产量最低但差异不显著(P＞0.05),其相对增重低于proE组和proC组.结果表明,proEC双抗原DNA疫苗对E.tenella攻虫表现出一定双抗原协同保护效应.     

中国部分地区绵羊肺炎支原体的基因多态性分析

为了解绵羊肺炎支原体(Mycoplasma ovi pneumoniae,Mo)在我国部分地区的流行病学特征,采用随机扩增多态性DNA(RAPD)及限制性片段长度多态性(RFLP)方法对26株Mo基因多态性进行了研究,并利用NTsys2.10e软件对获得的多态性图谱进行了聚类分析.结果在相似系数为0.70时,26株Mo可分成6个RAPD群或6个RFLP群;相似系数为0.90时,分成18个RAPD群或18个RFLP群;相似系数为1.00时,分成25个RAPD群或26个RFLP群.结果表明,我国Mo存在高度的基因多态性,这种多态性与地域差异相关,与宿主来源也具有一定的相关性.研究结果为了解我国Mo的分子流行病学特征打下了基础,也为建立有效的Mo诊断方法和疫苗研制提供了参考依据.     

用RAPD标记分析部分柞蚕二化性品种资源的遗传多样性

采用RAPD标记技术,对36份柞蚕(Antheraea pernyi)二化性品种资源的遗传多样性进行分析。PCR扩增及琼脂糖凝胶电泳结果表明:利用筛选出的19个随机引物在供试材料中共检测到123个信息位点,其中多态性位点114个,多态性比率92.68%;每一个引物可检测出3～9个位点,其多态性比率不尽相同,平均扩增出6.47条带;各品种具有不同数量的特异性带型。供试品种资源的Nei氏遗传多样性指数(h)为0.324 3±0.151 5,Shannon信息指数(I)为0.487 1±0.197 5,表明长期同地保存的二化性柞蚕品种资源同样具有丰富的遗传多样性;供试品种的Nei氏遗传一致性范围在0.495 9～0.837 4之间,达0.6以上的占96.59%,表明多数品种资源的相似程度较高。36份品种资源分别以辽宁省蚕业科学研究所育成种为主和以国内外引进种为主聚为两大类群。     

湖南省鸡蛔虫线粒体nad1基因部分序列的测定及分析

本研究旨在阐明鸡蛔虫湖南分离株线粒体烟酰胺腺嘌呤二核苷酸(NADH)脱氢酶亚单位1(nad1)基因部分序列(pnad1)遗传变异情况。利用聚合酶链式反应(PCR)扩增鸡蛔虫的pnad1,应用Clustal X 1.81程序对序列进行比对。结果显示,所获得的pnad1序列长度一致,为408 bp。由于鸡蛔虫pnad1序列种内相对保守,种间差异较大,故可作为种间遗传变异研究的标记,从而为鸡蛔虫的分类鉴定及进一步的分子流行病学调查奠定了基础。