Isolation and Characterization of Pathogenic Listonella anguillarum of Diseased Haft-Smooth Tongue Sole （Cynoglossus semilaevis Günther）     

TANG Xiaoqian ZHOU Li ZHAN Wenbin 《青岛海洋大学学报(英文版)》2008,(3):343-351

Studies were conducted to determine the cause of the acute mortality of half-smooth tongue sole Cynoglossus semilaevis Günther juveniles in a fish farm in Jimo, Shandong Province, China, in June 2006. Gross signs of the diseased tongue sole included several petechiae and ecchymoses on the body and fin necrosis and hemorrhagic lesion at the base of the fin. Bacteria were isolated from kidney, liver and hemorrhagic lesions of the diseased tongue sole. Among 14 strains, SJ060621 was proved to be highly virulent to juvenile tongue sole with LD50 value of 〈1.0×10^5 colony forming units （CFU）mL^-1, while the remaining 13 were avirulent. Among the 16 antibiotics tested, SJ060621 was sensitive to gentamicin and nitrofurantoin. It was identified as Listonella anguillantm with conventional plate and tube tests in combination with API 20E analysis. 16S rRNA gene and partial HSP60 gene sequenceing analysis revealed that the strain was highly homologous with L. anguillarum. Examination of the infected musculature by electron microscopy indicated numerous bacteria and lots of macrophages containing phagocytosed bacteria. Histopathological investigations revealed severe necrotic degenerative changes in the infected organs. Indirect immunofluorescence assay （IFA） was employed to detect the location of occurrence of bacteria, and bacteria were found in aggregations in the inflammatory areas in musculature.  相似文献   

泥蚶(Tegillarca granosa) cDNA 文库的构建及铁结合蛋白基因(Ferritin)序列分析     

贺静静  李晔  李太武  苏秀榕  王孟前  李松伟  周军  马斌  李强芳 《海洋与湖沼》2009,40(3):289-295

取正常泥蚶肌肉组织获得总RNA,纯化mRNA后,利用含SfiⅠ酶切位点的CDSⅢ引物逆转录合成cDNA第一链,通过LD—PCR合成cDNA双链,经胶回收除去短片段,与pDNR—LIB载体进行连接,电转化到大肠杆菌DH10B中,构建形成原始文库,进行滴度测试和重组率鉴定。结果表明原始文库的滴度为3．17×10^5cfu／ml,重组率为86．3％,插入片段多在500--2500bp间。随机挑选458个克隆测序,经分析获得94种已知基因及87种未知基因。其中包括铁结合蛋白（Ferritin）的全长cDNA序列。该基因序列全长895bp,5’-端非编码区为163bp,3’-非编码区为213bp,开放阅读框为519bp,编码172个氨基酸。推测其蛋白分子量和等电点分别为20kDa和4．89。氨基酸序列与皱纹盘鲍、太平洋牡蛎、血红扇头蜱、文昌鱼、中华蟾蜍、非洲爪蟾、小家鼠以及人等的同源性有62％-82％。比对结果表明,铁结合蛋白基因在动物进化中高度保守。  相似文献   

一株甲胺磷高效降解菌——巨大芽孢杆菌(Bacillus megaterium)的分离及其分子鉴定   总被引：2，自引：0，他引：2  

刘文海  邓先余  向言词  邱山红 《海洋与湖沼》2009,40(2):170-175

湘潭南天化工厂的甲胺磷废水未经处理直接排放,对湘江及下游的湖泊造成了一定的污染,利用解磷微生物去除江河湖泊中的甲胺磷污染是一条有效的途径.本文作者从被该厂甲胺磷废水污染的湖泊中分离细菌样品,以甲胺磷为唯一碳源和能源,经过定向筛选,得到一株可高效降解甲胺磷的菌株HN001.气相色谱测定结果表明,此菌株对甲胺磷的降解率在48h和96h分别为49.24%和98.20%.对其进行了常规生理生化测试,结果表明,该菌株与巨大芽孢杆菌的表型特征非常相似.为了进一步确定HN001的分类学地位,测定了其16S rRNA基因序列,分析了相关细菌相应序列的同源性,构建了分子系统发生树,结果表明菌株HN001与巨大芽孢杆菌的亲缘关系最近.综合上述结果,菌株HN001可鉴定为巨大芽孢杆菌(Bacillus megaterium).  相似文献   

中华鳖（Pelodiscus sinensis）不同种群酪氨酸酶（TYR）基因的克隆及其多态性分析   总被引：1，自引：0，他引：1  

张永正  张海琪  黄利英  丁诗华 《海洋与湖沼》2009,(3)

以中华鳖4个种群80个个体（太湖种群、日本品系种群、台湾引进种群和“清溪乌鳖”种群）肌肉基因组DNA为模板,利用已知酪氨酸酶（Tyrosinase,TYR）基因部分序列设计合成特异引物进行PCR扩增,克隆并测定了TYR基因的核苷酸序列。扩增所得的696bp序列中共有13个多态性核苷酸位点。中华鳖体色变异种群的3个个体均在第186核苷酸位点出现相同的变异。以此设计酶切位点,选用SmaⅠ内切酶进行RFLP,结果显示80个中华鳖个体的TYR基因存在多态性。AA型基因除在中华鳖体色变异种群内未发现外,其余三个种群内的频率大小顺序为日本品系种群（60％）〉台湾引进种群（30％）〉太湖种群（20％）,以省外品种为丰富;AB基因型频率大小顺序为“清溪乌鳖”种群（70％）〉台湾引进种群（50％）〉太湖种群（40％）〉日本品系种群（30％）。  相似文献   

Cytogenetic characterization of olive flounder Paralichthys olivaceus: DNA content, karyotype, AgNORs and location of major ribosomal genes     

WANG Xubo  ZHANG Quanqi  CHEN Yanjie  QI Jie  WANG Zhigang  WANG Xinglian 《海洋学报(英文版)》2009,28(4):72-77

A cytogenetic analysis of Paralichthys olivaceus was carried out using the flow cytometry method for DNA content, silver staining for the nucleolus organizer region (AgNORs) identification and one-color fluorescence in situ hybridization (FISH) for chromosomal mapping of major ribosomal genes. Nuclear DNA content was estimated by flow cytometry method using Gallus domesticus erythrocytes as the internal reference standard. The C-value of this species was (0.737±0.024) pg, and the DNA contents of each chromo...  相似文献   

Cloning and Analysis of Calmodulin Gene from Porphyra yezoensis Ueda （Bangiales, Rhodophyta）   总被引：2，自引：0，他引：2  

Mengqiang Wang  Yunxiang Mao  Yunyun Zhuang  Fanna Kong  Zhenghong Sui 《中国海洋大学学报(英文版)》2009,8(3):247-253

In order to understand the mechanisms of signal transduction and anti-desiccation mechanisms of Porphyra yezoensis, cDNA and its genomic sequence of Calmodulin gene (CaM) was cloned by the technique of polymerase chain reaction (PCR) based on the analysis of P. yezoensis ESTs from dbEST database. The result shows that the full-length cDNA of CaM consists of 603 bps including an ORF encoding for 151 amino acids and a terminate codon UGA, while the length of genomic sequence is 1231 bps including 2 exons and 1 intron. The average GC content of the coding region is 58.77%, while the GC content of the third position of this gene is as high as 82.23%. Four Ca²⁺ binding sites (EF-hand) are found in this gene. The predicted molecular mass of the deduced peptide is 16688.72 Da and the pI is 4.222. By aligning with known CaM genes, the similarity of CaM gene sequence with homologous genes in Chlamydomonas incerta and Chlamydomonas reinhardtii is 72.7% and 72.2% respectively, and the similarity of the deduced amino acid sequence of CaM gene with homologous genes in C. incerta and C. reinhardtii are both 71.5%. This is the first report on CaM from a species of Rhodophyta.  相似文献   

16S rRNA Gene Phylogenesis of Culturable Predominant Bacteria from Diseased Aposlichopus japonicus （Holothuroidea, Echinodermata）     

MA Haiyan JIANG Guoliang WU Zhiqiang WANG Xin 《中国海洋大学学报(英文版)》2009,8(2)

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深海微生物极端酶的研究进展     

李升康  王玉桥 《海洋科学》2009,33(5):98-102

21世纪是海洋的世纪,海洋生物资源的开发和利用已成为世界各海洋大国竞争的焦点之一,其中基因资源的争夺更是焦点中的重点.  相似文献   

花鲈PPARα基因实时荧光定量PCR质粒标准品的构建     

方雯  钱云霞 《台湾海峡》2009,28(4):488-491

利用SYBR荧光定量PCR技术,制备用于花鲈（Lateolabrax japonicus）PPARα基因实时荧光定量PCR检测的质粒标准品．通过PCR扩增出目的片断344-bp,纯化后连接至pMDI8-T载体,转化宿主菌E．coli DH5α．通过PCR鉴定、双酶切鉴定和测序分析后抽提重组质粒,测定其拷贝浓度,10倍梯度稀释成标准品模板10^8～10^4 copy／mm^3,进行荧光定量PCR分析．结果显示,循环阈值（Ct）与起始模板量的对数值之间有着良好的线性关系．  相似文献   

Cloning and Characterization of Largemouth Bass （Micropterus salmoides） Myostatin Encoding Gene and Its Promoter   总被引：1，自引：0，他引：1  

LI Shengjie BAI Junjie WANG Lin 《中国海洋大学学报(英文版)》2008,7(3)

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