山东近海褐牙鲆自然与养殖群体生化遗传结构及其遗传变异的比较分析

山东近海褐牙鲆自然群体活样本共 79尾 ,分别于 1 996年 5月、1 997年 1月和 1 998年 4月采自青岛近海 ;养殖群体活样本 5 2尾于 1 997年 1 2月采自山东荣成寻山养鱼场。采用水平淀粉胶和垂直聚丙烯酰胺凝胶电泳方法及 3种缓冲系统 (TC、EBT和TG)分别对自然和养殖群体的 1 5种同工酶进行生化遗传分析。结果表明 ,山东近海牙鲆自然群体的多态基因座位比例 ( 31 .0 % )和群体平均杂合度 ( 0 .0 80 2 )都明显高于养殖群体 ( 2 4 .1 % ,0 .0 788) ;在自然群体的 9个多态基因座位、养殖群体的 7个多态基因座位中 ,除了Cat(P <0 .0 5 )和Idhp - 1(P <0 .0 5 ,养殖群体中 )有显著差异、Ldh -C(P <0 .0 1 )完全偏离Hardy -Weinberg定律外 ,其余多态座位基因频率均符合Hardy-Weinberg遗传平衡定律     

中国对虾输精管结构及精子形成

采用激光扫描共聚集显微镜及电子显微镜技术，对中国对虾输精管结构及精子的形成过程进行研究。结果表明，中国对虾的输精管可分为近端、中间、远端输精管及壶腹4部分，输精管壁各部分的基本结构相似，但分泌结构的结构有较大的差别。输精管自中间膨大部开始，管腔逐渐被一隔膜分为大小不等的两部分，较大的腔内充满了很多精子，输精管较小的腔内充满胶体状物质，中国对虾精子细胞在精巢内产生，在两条输精管内逐渐发育，经过细胞质的分化并与精子外部进行物质的交换，形成顶体、亚顶体及棘突。核的变化则经历了染色质的解凝和核膜的消失，最后形成成熟的精子。     

栉孔扇贝NDPK 基因的BAC-FISH 定位研究

栉孔扇贝(Chlamys farreri)是我国北部沿海一个非常重要的养殖种类。核苷二磷酸激酶(NDPK)是一类广泛存在、高度保守,在细胞能量和信息传递中具有重要作用的酶。作者利用BAC-FISH技术将两个包含栉孔扇贝NDPK基因的BAC克隆定位到染色体同一位置上,该结果验证了物理图谱相关contig组装的可靠性,同时NDPK基因的染色体定位将对深入研究该基因的结构、功能以及应用于生产实践提供理论支撑。本研究结果将对栉孔扇贝染色体的深入研究以及染色体鉴别、图谱整合等工作提供重要参考。     

昆虫围食膜结构与功能概述及其对对虾WSSV 病防治研究的启示

围食膜(Peritrophic matrix,PM)是无脊椎动物肠道内包围摄入食物的一种特殊结构,是由几丁质纤维网在黏附于其上的蛋白质(围食膜蛋白)相互作用下形成的半通透性膜状结构[1]。已发现的围食膜有两种类型,即I型和II型[2]。I型围食膜是由中肠上皮细胞分泌并覆盖于整个中肠表面;II型则由贲门附近的细胞分泌,随着取食产生,包在食物表面并随着消化废物排出体外[3]。类似于脊椎动物消化道的黏液层,它是中肠上皮细胞和食物之间的一个物理屏     

Rapid,sensitive detection of Vibrio anguillarum using loop-mediated isothermal amplification

Vibrio anguillarum is an important bacterial pathogen of aquatic organisms and a significant problem in aquatic farming.The rapid detection and identification of V.anguillarum,and other pathogens that infect marine organisms,is crucial to effective disease management.In this study,we developed a loop-mediated amplification (LAMP) assay to detect V.anguillarum in an hour in a single tube without the need for thermal cycling.Conserved regions of the metalloproteinase (empA) gene of V.anguillarum served as the targets for primer design.A fragment of the empA gene was amplified at 65°C in the presence of the primer mixture and Bst DNA polymerase.In the optimized LAMP assay,6.7 pg of V.anguillarum DNA could be detected.Six strains of V.anguillarum and 17 strains of non-V.anguillarum bacteria were used in this study to evaluate the species specificity of the primers.The six V.anguillarum strains gave a positive result in the LAMP assay.This method was also validated in V.anguillarum-infected fish.This LAMP method is more sensitive than PCR in the detection of V.anguillarum and shows good species specificity.The LAMP assay is therefore an effective method for the quick detection of V.anguillarum both in the laboratory and in the field.     

Proteomic analysis of acute responses to copper sulfate stress in larvae of the brine shrimp, Artemia sinica

Proteomics was used to reveal the differential protein expression profiles of acute responses to copper sulfate exposure in larvae of Artemia sinica.Fourteen differentially displayed protein spots were detected and seven of them were identified.Three spots were up-expressed and identified:actin, heat shock protein 70,and chaperone subunit 1;three down-regulated proteins were identified:arginine kinase,elongation factor-2,and glycine-rich protein;and a newly expressed protein was identified as peroxiredoxin.The study indicates the involvement of all the differentially expressed proteins in the early responses of protein expression,and in the survival of A.sinica in the presence of copper and other heavy metals;the findings improve understanding of the organism’s adaptive responses and resistance.     

Chromosomal localization of 5S rDNA in Chinese shrimp (Fenneropenaeus chinensis): a chromosome-specific marker for chromosome identification

Chinese shrimp (Fenneropenaeus chinensis) is an economically important aquaculture species in China. However, cytogenetic and genomic data is limited in the organism partly because the chromosomes are difficult to isolate and analyze. In this study, fluorescence in-situ hybridization (FISH) was used to identify the chromosomes of F. chinensis. The 5S ribosomal RNA gene (rDNA) of F. chinensis was isolated, cloned and then used as a hybridization probe. The results show that the 5S rDNA was located on one pair of homologous chromosomes in F. chinensis. In addition, triploid shrimp were used to evaluate the feasibility of chromosome identification using FISH and to validate the method. It was confirmed that 5S rDNA can be used as a chromosome-specific probe for chromosome identification in F. chinensis. The successful application of FISH in F. chinensis shows that chromosome-specific probes can be developed and this finding will facilitate further research on the chromosomes of penaeid shrimps.     

中国对虾6种组织cDNA文库的构建

以中国对虾血液、眼柄、卵巢、雌虾头胸部、雄虾头胸部和三倍体对虾头胸部组织为材料,采用异硫氰酸胍-酸酚法提取总RNA;用磁珠法纯化mRNA;用Uni ZAP cDNA合成试剂盒合成双链cDNA并进行修饰,将cDNA定向连接在UniZAP载体上;经Gigapack Gold Package包装试剂盒包装成为噬菌体颗粒,转染宿主XL1 Blue MRF’菌株细胞,形成初级文库.初级文库经转染进一步扩增,形成稳定的cDNA文库.6个文库的库容在0.2×106~1.3×106之间,重组率都超过90%.从各文库随机取出6~10个清晰的噬菌斑进行PCR扩增,用琼脂糖凝胶电泳检测,其插入片段长度为500~2500bp.由初步功能基因克隆获得阳性结果.多项指标表明,所构建的对虾cDNA文库质量较高,为进一步筛选目的基因、EST测序和制作基因芯片提供了有效的工具.     

热休克蛋白研究进展

热休克蛋白是生物体在不利环境因素刺激下应激合成的一族进化上高度保守的蛋白质。综述了热休克蛋白在热休克因子和热休克元件调节下的基因表达调控,并讨论了热休克蛋白的生物学功能和它们在海洋生物养殖方面潜在的应用意义。