四色荧光标记DNA自动测序中一例典型错读码的校正   总被引：3，自引：0，他引：3  

马洪明  路新枝  王勇 《海洋科学》2002,26(3):10-13

ＤＮＡ测序是现代分子生物学研究中的重要技术。随着人类基因组计划的实施 ,ＤＮＡ测序技术虽然在原理上没有新突破 ,仍然依赖于Ｓａｎｇｅｒ的酶法 ,但在不断提高自动化程度、降低成本方面取得了很大的进展 ,测序效率目前已有很大提高。大规模测序一个碱基的成本从１９８６年的０．５美元降到目前的约７美分。目前各种ＤＮＡ自动测序技术都是在荧光标记技术的基础上发展起来的。采用荧光标记的自动测序仪已在各个领域被广泛应用。四色荧光标记自动测序采用４种具有不同发射波长的荧光染料标记由聚合链终止反应产生的一系列终止片段 ,经过聚…  相似文献   

白斑综合症病毒(WSSV)的宿主调查   总被引：24，自引：3，他引：24  

雷质文  黄倢  史成银  张立敬  俞开康 《海洋与湖沼》2002,33(3):250-258

用地高辛 (DIG)标记的WSSVDNA探针斑点杂交与原位杂交技术 ,在中国对虾、斑节对虾、南美白对虾、刀额新对虾、脊尾白虾、天津厚蟹、日本大眼蟹体内检测到了WSSV ,它们是WSSV的天然宿主 ;在经人工感染的哈氏美人虾、短脊鼓虾、克氏原螯虾、肉球近方蟹、滕壶体内检测到了WSSV ;在球形侧腕水母、病虾池的桡足类等浮游生物、卤虫无节幼体以及人工浸泡感染卤虫成体体内没有检测到WSSV。经原位杂交检测 ,虾类的甲壳下上皮、胃上皮、附肢、造血组织、鳃等组织器官均可被WSSV侵染 ,其中甲壳下上皮和鳃对WSSV敏感 ;蟹类的甲壳下上皮和鳃对WSSV敏感 ;在中国对虾、南美白对虾、脊尾白虾、注射感染的克氏原螯虾的精巢中 ,精荚的结缔组织细胞和血细胞呈阳性 ,在中国对虾、脊尾白虾以及注射感染的短脊鼓虾的卵巢中 ,结缔组织细胞和滤泡细胞被WSSV感染。  相似文献   

利用RAPD和AFLP标记初步构建太平洋牡蛎的遗传连锁图谱   总被引：24，自引：0，他引：24       下载免费PDF全文

李莉  郭希明 《海洋与湖沼》2003,34(5):541-551

利用RAPD和AFLP标记 ,以一回交家系 [(Miyagi×Hiroshima)×Miyagi]为作图群体 ,构建了太平洋牡蛎的遗传连锁图谱。用经过筛选的 3 3个RAPD引物和 1 1个AFLP引物组合 ,对父母本和 80个子代个体进行了遗传分析。共得到母本分离标记 1 93个 ,其中 1 44个符合1∶1孟德尔分离规律。父本分离标记 1 5 6个 ,其中 1 1 1个符合 1∶1孟德尔分离规律。雌性框架图包括 99个遗传标记 ,定位在 1 2个连锁群中 ,覆盖 985 2cM ,标记间平均间隔 1 1 3cM。另外有 3个三联体 ,7个连锁对 ,图谱共覆盖 1 1 6 5 7cM。雄性框架图包括 72个遗传标记 ,分布在 8个连锁群 ,覆盖 81 1cM ,标记间平均间隔 1 2 7cM。另外有 4个三联体 ,3个连锁对 ,图谱共覆盖93 1 8cM。  相似文献   

刀额新对虾染色体核型及细胞核DNA含量   总被引：9，自引：0，他引：9  

张晓军  周岭华  相建海 《海洋与湖沼》2002,33(3):225-231

以刀额新对虾胚胎、无节幼体、卵巢、精巢等为材料 ,空气干燥法制备染色体 ,并初步进行核型分析。结果表明 ,刀额新对虾的染色体数目为 2n=78,n =39,核型为 2n=78=40M 1 0SM 1 4ST 1 4T。以刀额新对虾血淋巴、卵巢、肌肉、鳃为材料 ,以鸡血细胞为DNA标准 (2 50pg/ 2c) ,使用PartecCCAⅡ型流式细胞仪测定了刀额新对虾细胞的基因组DNA含量 ,其值为鸡血细胞的 1 75倍 ,绝对含量为 4 375pg/ 2c。  相似文献   

Effect of strong winds on the nutritional condition of anchovy (Engraulis encrasicolus L.) larvae in the Bay of Biscay, Northeast Atlantic, as inferred from an early field application of the DNA/C index     

Bergeron  Jean-Pierre 《ICES Journal of Marine Science》2000,57(2):249-255

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中国花鲈细胞色素b基因片段的序列研究(英文)     

高天翔  张秀梅  陈大刚  张美昭  任一平  张亚平 《中国海洋大学学报(自然科学版)》2001,31(2)

参考鳗鲡等鱼类线粒体 DNA序列进行了中国花鲈线粒体 DNA细胞色素 b基因片断的引物设计、PCR扩增及其序列测定。得到中国花鲈的碱基序列为 4 10 bp,其 A、T、G、C含量分别为 10 1bp(2 4 .6 3% )、112 bp(2 7.32 % )、72 bp(17.56 % )、12 5bp(30 .4 9% ) ,与鳗鲡等其他鱼类相同基因片断序列碱基含量相似。  相似文献   

Application of the first internal transcribed spacer(ITS-1) of ribosomal DNA as a molecular marker to population analysis in farrer''s scallop Chlamys farreri     

YU Ziniu  WEI Xiaohu  KONG Xiaoyu  YU Shanshan 《海洋学报(英文版)》2007,26(1):93-100

Sequence variation of the first internal transcribed spacer of ribosomal DNA （ ITS - 1 ） was examined and its application to the study of genetic variation was explored in four populations of farter＇ s scallop Chlamys farreri. ITS - 1 fragments, with a length of about 300 bp,of 78 individuals collected from Dalian, Qingdao, Yantai in China and Korea respectively were amplified via PCR, cloned and sequenced. Intra-genomic variation was examined by sequencing several clones of single individuals. Alignment and polymorphism analysis detected 44 haplotypes and 50 polymorphic sites which consist of 30 substitutions and 20 indels, indicating a high level of polymorphisms. Sequence analysis also showed a very low level of intra-individual variation. All these features validated the feasibility of application of ITS - 1 fragment to population analysis. Polymorphism analysis showed that the Korea sample has the richest genetic variation, followed by Yantai and Qingdao samples. AMOVA （analysis of molecular variance） showed that the majority （96.26%） of genetic variation was distributed within populations and 3.74% resulted from among populations, but with P 〈 0.05 （ = 0.042）, indicating that the populations in this study have significant divergence. This output was basically concordant with the result arising from RAPD data and different from that from mitochondrial 16S rDNA sequence data. Discussion on this inconsistency was made accordingly.  相似文献   

3种蛏类线粒体16SrRNA和COI基因片段的序列比较及其系统学初步研究   总被引：15，自引：0，他引：15  

陈丽梅  孔晓瑜  喻子牛  于珊珊  徐晖 《海洋科学》2005,29(8):27-32

对3种蛏类大竹蛏（Solen grandis），长竹蛏（Solen strictus）和小刀蛏（Cultellus attenuatus）的线粒体16SrRNA和COI基因片段序列进行了比较并对其系统学进行了初步研究。得到的序列总长度分别为472-481bp（16S）和658bp（COI）。3种蛏序列的碱基组成均显示出较高的A＋T比例（16SrRNA基因62．1％；COI基因62．8％）。对位排序比较表明，16SrRNA片段种内个体间变异较小，3种类间存在128个碱基变异位点（其中包括127个简约信息位点）和5个插入／缺失位点，总共12个碱基长；COI片段有200个碱基存在变异，其中包括191个简约信息位点，不存在任何插入／缺失位点。数据分析结果表明16SrRNA和COI基因片段大竹蛏与长竹蛏两片段的遗传距离分别为0．0856和0．1712，两竹蛏类与小刀蛏的遗传距离分别为0．3054，0．2798和0．2662，0．2933。作者认为小刀蛏与竹蛏之间的遗传距离已达到科之间的水平，结果支持将其提升为刀蛏科的分类观点。3种蛏类线粒体16SrRNA和COI基因在种间明显的多态性，证实了16SrRNA和COI基因序列均普遍适用于蛏类种及以上阶元的系统学分析。  相似文献   

Detection of DNA damage in mussels and sea urchins exposed to crude oil using comet assay     

Taban IC  Bechmann RK  Torgrimsen S  Baussant T  Sanni S 《Marine environmental research》2004,58(2-5):701-705

The single-cell microgel electrophoresis assay or the comet assay was used to evaluate DNA damage of dispersed crude oil on sea urchins (Strongylocentrotus droebachiensis) and mussels (Mytilus edulis L.). Sea urchins were exposed to 0.06 and 0.25 mg/L dispersed crude oil in a continuous flow system, while the mussels were exposed to 0.015, 0.06 and 0.25 mg/L dispersed crude oil. Sea urchin coelomocytes and mussel haemocytes were sampled after 4 and 5 weeks exposure, respectively. In the sea urchin coelomocytes, there was a significant concentration-related increase in the percentage of DNA in comet tail. In mussel haemocytes, there was a significantly higher percentage of DNA in comet tail for all treatments compared to the control. The responses were concentration-related up to 0.06 mg/L oil. The two highest exposure concentrations of mussels were not significantly different from each other. These results indicate that the comet assay can be used for biomonitoring of DNA damage in marine invertebrates following oil contamination.  相似文献   

Molecular identification of ascaridoid nematodes from the deep-sea onion-eye grenadier (Macrourus berglax) from the East Greenland Sea   总被引：2，自引：0，他引：2  

Esra Kellermanns  Sven Klimpel  Harry Wilhelm Palm 《Deep Sea Research Part I: Oceanographic Research Papers》2007,54(12):2194-2202

Macrourus berglax from the East Greenland Sea was studied for the presence of ascaridoid nematodes in 2001, 2002 and 2003. The fishes were collected between 278 and 413 m water depth using a benthopelagic net. Based on the amplification of the internal transcribed spacer ITS-1, 5.8S, ITS-2 and flanking sequences (=ITS+), three ascaridoid nematode species were identified. The prevalence of infestation during the 3 years ranged from 42.9% to 62.9% and 22.9% to 40.0% for the anisakids Anisakis simplex (s.s.) and Pseudoterranova decipiens (s.s.), respectively, and from 28.6% to 60.0% for the raphidascarid Hysterothylacium aduncum. A total of 18 specimens, two of each species and examination year, revealed no sibling species, suggesting a limited distribution of other ascaridoid siblings into the deep sea. The ITS-1, 5.8S and ITS-2 sequences of A. simplex (s.s.) from the East Greenland Sea did not differ from previously published sequence data (GenBank) from other regions in the Atlantic and Pacific oceans. The sequences of P. decipiens (s.s.) corresponded most closely to those of specimens from Richardson Bay, western Pacific, and differed in four positions (0.5%). They corresponded least to those of specimens from Japan (1.5%). The sequence data for H. aduncum differed in two positions in the ITS-1 (0.2%) and three positions in the ITS-2 (0.3%) from sequences from Japan. A high genetic similarity between the regions can be explained by (a) extensive final host migration in the case of A. simplex (s.s.), (b) an overlapping distribution of final host populations along the continental shelves for P. decipiens (s.s.) and (c) a low host specificity and large population size in the intermediate and final hosts for H. aduncum. The occurrence of the identified species in the macrourid fish underlines the potential of cosmopolitan ascaridoid nematodes to distribute not only horizontally but also vertically in the deep sea.  相似文献