蛤蜊科3种贝类16SrRNA基因片段及ITS2核苷酸序列分析

利用PCR技术分别扩增连云港及启东沿海蛤蜊科的西施舌（Coelomactra antiquata）、中国蛤蜊（Mactra chinensis）和四角蛤蜊（Mactra veneriformis）3种双壳贝的16SrRNA基因片段和ITS2核苷酸序列．测序后用DNAstar软件分析了核苷酸差异。结果显示：三种贝类16SrRNA基因片段长度相同，均为306bp（去除引物）．核苷酸存在多态性。共有45个变异位点，54个核苷酸发生了变异。全部为碱基置换。西施舌与中国蛤蜊此片段核苷酸的同源性为88．9％．与四角蛤蜊的同源性为88．6％．中国蛤蜊与四角蛤蜊的同源性为90．6％。三种蛤蜊ITS2序列分别为390bp（西施舌）、441bp（四角蛤蜊）和466bp（中国蛤蜊）。存在长度多态性．ITS2核苷酸差异分析结果显示．西施舌与中国蛤蜊的同源性为70．9％-71．1％，西施舌与四角蛤蜊的为70．5％-71．0％。中国蛤蜊与四角蛤蜊的同源性为88．1％-88．8％。ITS2序列分析结果与16SrRNA基因片段分析结果一致．2种分子分析法均显示中国蛤蜊与四角蛤蜊的亲缘关系近。     

盐胁迫对番茄同工酶基因表达的影响

分析了普通番茄（Lycopersicumesculentum）在NaCl胁迫下酯酶（EST）和过氧化物酶（PRX）同工酶的变化，考察了盐胁迫对10个同工酶位点21个等位基因在特定组织中表达的影响。发现许多EST和PRX等位基因在盐胁迫下表达，产生盐诱导同工酶；另有些等位基因的表达受盐抑制，它们在盐胁迫下的表达活性显著减弱或消失。实验表明，EST和PRX同工酶表达的变化与番茄对盐胁迫的遗传适应性有密切关系。     

Microbial Diversity in Nankai Trough Sediments at a Depth of 3,843 m

Dense populations of bivalves, primarily Calyptogena sp., were observed at cold seeps of the Nankai Trough. Bacterial input to the sediment was estimated through determination of phospholipid ester-linked fatty acid (PLFA) and DNA profiles. Results indicated a bacterial biomass of 10⁹ cells (g dry wt)^-1 while individual fatty acid profiles revealed a predominance of monounsaturated fatty acids, mainly 18:1 isomers. The presence of these fatty acids can be interpreted to reflect a response to low temperature and a predominance of psychrophilic bacteria. DNA fragments encoding bacterial ribosomal RNA small-subunit sequences (16S rDNA) were amplified by the polymerase chain reaction method using DNA extracted directly from the sediment samples. From the sequencing results, at least 19 kinds of bacterial 16S rDNAs related to mostly the Proteobacteria and a few gram-positive bacteria were identified. These results suggest that the bacterial community in the Nankai Trough sediments consists of mainly bacteria belonging to the Proteobacteria , , and subdivisions. Bacteria belonging to the and subdivisions, which are known to include epibiont and sulfate reducing bacteria, respectively, were mostly detected in the sediment obtained from inside the area of the Calyptogena community, and the -Proteobacteria may function to supply reduced sulfur to bacterial endosymbionts of Calyptogena.     

中华齿米虾胰蛋白酶基因的研究

以中华齿米虾(Neocaridina denticulata sinensis)基因组DNA为模板,根据胰蛋白酶基因保守序列设计简并引物,利用PCR技术获得一个基因。序列分析表明此基因与已报道的凡纳滨对虾(Litopenaeus vannamei)胰蛋白酶基因有较高的相似度,序列中包含一个内含子和两个不完整的外显子,编码182个氨基酸残基。该氨基酸序列与凡纳滨对虾胰蛋白酶氨基酸序列的相似度为83.5%;含有胰蛋白酶所特有的His、Asp和Ser组成的活性三联体、决定胰蛋白酶底物专一性的Asp、底物结合部位的G1y残基。综合分析认定该基因为胰蛋白酶基因片断。将其氨基酸序列与报道的其他动物胰蛋白酶氨基酸序列进行了系统进化分析。系统进化分析的结果与现有以表型特征为依据的虾分类结果是一致的。     

6-磷酸山梨醇脱氢酶(gutD)基因克隆、测序和表达

本研究根据6-磷酸山梨醇脱氢酶基因(gutD)两端序列设计引物，以假单胞杆菌(Pseudonmonas XM)的总DNA为模板通过PCR的方法克隆gutD基因并进行序列分析，将克隆得到的gutD基因与原核表达载体pBV220连接并转化大肠杆菌(Escherichia coli)JM101后检测其蛋白表达及菌株生长耐盐性。     

A cDNA microarray technique applied for analysis of global gene expression profiles in tributyltin-exposed ascidians

To analyze global gene expressions, we constructed a cDNA microarray from a basal chordate, the ascidian Ciona intestinalis. Ciona is a cosmopolitan species and a genomic analysis of Ciona revealed that ascidians had approximately 15,500 protein-coding genes. Our "Ciona intestinalis cDNA chip version 1 (Ci cDNA chip ver. 1)" has arrayed 13,400 unique Ciona cDNAs. To establish a detection system for gene expression profiles in wild ascidians using a cDNA microarray, we analyzed gene expressions in the whole body of Ciona adults after exposure to 100 nM tributyltin (TBT) for 24 h. In our preliminary array data using Ci cDNA chip ver. 1, we found more than 200 genes that showed strong differential expressions. These genes encoded proteins that were concerned with stress response, detoxification, oxidoreduction reaction, biosynthesis, and catabolism. This, the first large cDNA microarray of this animal, should facilitate analyses of global gene expressions following exposure to TBT.     

Diagnosis of iridovirus in large yellow croaker by PCR

A rapid and sensitive PCR-based method for the detection of the large yellow croaker iridovirus(LYCIV) is described,which involves the amplification of a 295 bp fragment of the LYCIV ATPase gene from DNA isolated from naturally infected fish spleen.Sequencing of LYCIV ATPase gene fragment showed it shared 100% nucleotide sequence homology with the corresponding region of the ATPase gene of red sea bream iridovirus(RSIV) and sea bass iridovirus(SBIV),suggesting that LYCIV was homologous with RSIV and SBIV at least in part of the gemone.The specificity and sensitivity of the PCR procedure were tested on the iridovirus-infected fishes,the expected fragment was detected from spleen DNA samples of infected fishes,whereas no fragments were amplified from healthy fish spleen DNA,white spot syndrome baculoviruses(WSBV) DNA and pseudorabies virus(PRV) DNA.Detection limit of this method was 10^-7 ng positive plasmid DNA containing target sequence,equal to about 100 virions.In the infected experiment,first positive detection(1/4) appeared at Day 3 post-infection,all fish(4/4) tested positive at Day 7,however obvious symptoms were observed at Day 8,so LYCIV infection could be detected prior to the appearance of obvious symptoms.These results indicate that this PCR method could be used for early,rapid and specific detection of LYCIV infection.     

耳鲍(Haliotis asinina)核糖体小亚基(18S rRNA)编码基因的克隆与序列分析

采用分子生物学的方法,对南海不同海区的两个地理群体耳鲍(Haliotis asinina)18S rRNA基因全长进行了克隆和序列分析,并将耳鲍18S rRNA基因的序列与NCBI数据库中已收录鲍的18SrRNA基因进行了比较。结果发现,南海耳鲍核糖体18S rRNA基因与耳鲍H.asinina isolate H11核糖体18S rRNA基因序列的相同率高达98%;同一地理群体内耳鲍核糖体18S rRNA基因序列完全一致;不同地理群体间耳鲍核糖体18S rRNA基因在碱基组成上的相似率为99%,仅在某些位点处发生了碱基替换,即腺嘌呤(T)被鸟嘌呤(G)替换;同时,将这两个不同群体中耳鲍的18S rRNA基因与泰国耳鲍18S rRNA基因序列进行比较分析发现,它们之间也只是发生了碱基替换。     

沉降监测多项式回归分析与神经网络预测

使用多项式和切比雪夫(Tchebyshev)多项式分别对沉降监测数据进行回归分析以预测未来沉降值,其中切比雪夫多项式的外推效果较好;应用前向BP神经网络对两种不同的单因子输入模式进行非线性函数逼近,并进行了不同采样步长的比较,实例表明将时间点作为网络的输入对沉降进行预测效果较好。