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Acta Oceanologica Sinica 2013年第2期32卷 52-58页

作者： SUN Yumiao LI Fuhua CHI Yanhong xiang jianhai Institute of Oceanology Chinese Academy of Sciences Graduate University Chinese Academy of Sciences

Recombinant bacteria secreting the transmembrane-truncated region of white spot syndrome virus （WSSV） envelope protein VP28 named DhpVB were constructed, using live Escherichia coli as a deliverer and a con- stitutive secretory expression plasmid as the expression vector. With the ability to deliver recombinant pro- tein products outside, DhpVB could make VP28 directly interact with the shrimp when injected into shrim- p. In order to test the protection potential, live DhpVB were injected into the marine shrimp Exopalamon carincauda Holthuis for three times and WSSV challenge was performed twice simultaneously at the last two injections of the bacteria. The results showed that DhpVB displayed statistically significant advantage over bacteria without VP28 after the second challenge with an average RI （Resistance index） of 0.67±0.08, compared with 0.41±0.09 of bacteria without VP28 （P 〈0.05）. The data suggested that a quasi-immune response may exist in E. carincauda and live bacteria carrying VP28 could enhance the shrimp resistance against WSSV.

关键词： WSSV VP28 Exopalamon carincauda live bacterial deliver

Heritability estimations of ammonia tolerance and survival of ridgetail white prawn Exopalaemon carinicauda

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Journal of Oceanology and Limnology 2022年第4期40卷 1662-1668页

作者： Ding LÜ Chengsong ZHANG Yang YU jianhai xiang Fuhua LI Key Laboratory of Experimental Marine Biology Institute of OceanologyChinese Academy of SciencesQingdao 266071China Laboratory for Marine Biology and Biotechnology Pilot National Laboratory for Marine Science and Technology(Qingdao)Qingdao 266237China Center for Ocean Mega-Science Chinese Academy of SciencesQingdao 266071China The Innovation of Seed Design Chinese Academy of SciencesWuhan 430072China

Ridgetail white prawn Exopalaemon carinicauda is an important economic marine species in *** improve the germplasm quality of cultured ***,for the fi rst time,we estimated the heritability of ammonia tolerance and survival of ***,covering 263 full-sib families in multiple *** models including binary linear animal model(BLA),binary linear sire-dam model(BLSD),probit threshold animal model(PTA),and probit threshold sire-dam model(PTSD)were *** estimated heritability of tolerance to ammonia in diff erent models ranged from 0.09 to 0.22,and that of survival rate ranged from 0.06 to *** the tolerance to ammonia,both the PTSD and BLSD models showed comparatively better prediction accuracies(0.48 and 0.47,respectively)than PTA and BLA models(0.40 and 0.26,respectively),while to the survival rate,all four models presented relatively low predictive abilities(0.16-0.26).This study provided important information for the future breeding program of ***.

关键词： heritability Exopalaemon carinicauda tolerance to ammonia survival rate

Foreign gene transfer into Chinese shrimps (Penaeus chinensis) with gene gun

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Chinese Science Bulletin 2001年第9期46卷 766-770,794页

作者： LIU Zhiyi xiang jianhai ZHOU Guoying GONG Zuxun Experimental Marine Biology Laboratory Institute of OceanologyChinese Academy of SciencesQingdao 266071China Shanghai Institute of Biochemistry Chinese Academy of SciencesShanghai 200031China

Plasmids pG DNA-RZ1 with a GFP (green fluorescent protein) reporter gene and a ribozyme gene incising penaeid white spot baculovirus (WSBV) were first introduced into the fertilized eggs of Chinese shrimps by gene gun. The treated and control samples of different development stages were observed with a fluorescent microscope. The transient expression of GFP gene was high in nauplius and zoea larvae. Results from RT-PCR and PCR for adults showed that the foreign genes had been transferred into the shrimps and had expressed the corresponding proteins. This work has established a transgenic method for penaeid shrimps, which will set base for the application of genetic engineering breeding into industry.

关键词： gene gun GFP transgenic shrimps.

Molecular cloning and expression analysis of Crustin-like gene from Chinese shrimp Fenneropenaeus chinensis

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Acta Oceanologica Sinica 2008年第2期27卷 81-92页

作者： LIU Fengsong LI Fuhua xiang jianhai DONG Bo LIU Yichen ZHANG Xiaojun ZHANG Liusuo College of Life Science Hebei University Baoding 071002 China Experimental Marine Biology Laboratory Institute of Oceanology Chinese Academy of Sciences Qingdao 266071 China

A new member of antimicrobial protein genes of the Crustin family was cloned from haemocytes of the Chinese shrimp Fennero- penaeus chinensis by 3 ′and 5′ RACE. The full-length cDNA of Crustin-like gene contains a 390 bp open reading frame, encoding 130 amino acids. The deduced peptide contains a putative signal peptide of 17 amino acids and mature peptide of 113 amino acids. The molecular mass of the deduced mature peptide is 12. 3 ku. It is highly cationic with a theoretical isoelectric point of 8.5. The deduced amino acids sequence of this Crustin showed high homology with those of Penaeus （ Litopenaeus ） setferus. Northern blotting showed that the cloned Crustin gene was mainly expressed in haemocytes, gill, intestine, and RNA in situ hybridization indicated that the Crustin gene was constitutively expressed exclusively in haemocytes of these tissues. Capillary elec- trophoresis RT-PCR analysis showed that Crustin was up-regulated dramatically from 12 to 48 h after a brief decrease of mRNA during first 6 h in response to microbe infection. The level of Crustin mRNA began to restore at 72 h post-challenge. This indicated that Crustin gene might play an important role when shrimps are infected by bacterial pathogen.

关键词： Crustin-like gene Fenneropenaeius chinensis molecular cloning expression analysis microbe challenge

Neuroanatomy and morphological diversity of brain cells from adult crayfish Cherax quadricarinatus

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Journal of Oceanology and Limnology 2018年第6期36卷 2368-2378页

作者： DUAN Hu JIN Songjun LI Fuhua ZHANG Xiaojun xiang jianhai Key Laboratory of Experimental Marine Biology Institute of OceanologyChinese Academy of Sciences Laboratory for Marine Biology and Biotechnology Qingdao National Laboratory for Marine Science and Technology

As in vertebrates, brains play key roles in rhythmic regulation, neuronal maintenance, diff erentiation and function, and control of the release of hormones in arthropods. But the structure and functional domains of the brain are still not very clear in crustaceans. In the present study, we reveal the structural details of the brain in the redclaw crayfish using hematoxylin-eosin staining and microscopic examination, firstly. The brain of crayfish is consist of three main parts, namely, protocerebrum, deutocerebrum, and tritocerebrum, including some tracts and commissures, briefly. Secondly, at least 9 kinds of brain cells were identified on the basis of topology and cell shapes, as well as antibody labeling. We also provide morphological details of most cell types, which were previously un-described. In general, four types of glia and three types of neurosecretory cells were described except cluster 9/11 and cluster 10 cells. Glia were categorized into another three main kinds:(1) surface glia;(2) cortex glia; and(3) neuropile glia in addition to astrocytes identified by GFAP labelling. And neurosecretory cells were categorized into I, Ⅱ and III types based on morphological observation. Finally, cluster 9/11 and 10 cells derived from the brain of crayfish, could be used for primary culture about 7–9 d under the optimized conditions. There results provide a resource for improving the knowledge of the still incompletely defined neuroendocrinology of this species. Using the crayfish as an animal model, we are easy to carry out further research in manipulating their endocrine system, exploring cellular and synaptic mechanisms so much as larval production on a small scale, such as in a cell or tissue.

关键词： Cherax quadricarinatus brain’s structure brain cells primary culture

Detection of the End Point Temperature of Thermal Denatured Protein in Fish and Chicken Meat Through SDS-PAGE Electrophoresis

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Journal of Ocean University of China 2009年第1期8卷 95-99页

作者： GAO Hongwei MAO Mao LIANG Chengzhu LIN Chao xiang jianhai ）Institute of Oceanology Chinese Academy of Sciences Qingdao 266003 P. R. China College of Food Science and Engineering Ocean University of China Qingdao 266003 P. R. China Shandong Entry-Exit Inspection and Quarantine Bureau of P. R. China Qingdao 266003 P. R. China

Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) was applied in the detection of the end point temperature (EPT) of thermal denatured protein in fish and meat in this study. It was also used in studying the thermal denatured temperature range of proteins in salmon and chicken meat. The results show that the temperature ranges of denatured proteins were from 65 ℃ to 75 ℃ , and these temperature ranges were influenced by the processing methods. Through SDS-PAGE, the features of repeated heating thermal denatured proteins under the same temperature and processing time were studied. The electrophoresis patterns of thermal denatured proteins determined through repeated heating at the same temperature did not exhibit any change. For the detection of cooked fish and meat samples, they were subjected to applying the SDS-PAGE method, which revealed an EPT ranging from 60 ℃ to 80 ℃ .

关键词： end point temperature detection SDS-PAGE electrophoresis

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中国水产种业发展过程回顾、现状与展望

中国农业科技导报 2013年第6期15卷 1-7页

作者：相建海中国科学院海洋研究所山东青岛266071

近四十年来,我国水产养殖业取得了长足的发展,并成为大农业中发展最快的产业之一。苗种是水产增养殖产业的源头和必要的物质基础,种质苗种是水产养殖业的核心问题。回顾了我国年轻的水产种业发展历程:最早期的养殖苗种来自于野生种群,... 详细信息

近四十年来,我国水产养殖业取得了长足的发展,并成为大农业中发展最快的产业之一。苗种是水产增养殖产业的源头和必要的物质基础,种质苗种是水产养殖业的核心问题。回顾了我国年轻的水产种业发展历程:最早期的养殖苗种来自于野生种群,及到人类驯化了亲本鱼,在人工条件下成功实施繁育,开始规模化育苗,才有了真正意义上的苗种产业;而良种的引入可以加快新产业形成。人工育种是在某一物种业已形成大规模人工繁育的基础上,特别是可以实现全人工繁殖的条件下,按照人们的意愿和方向开展的代代相继、不断选优的选育实践。20世纪90年代以来,我国水产特别是海洋水产种业受到重视,走向良性发展,取得了引人注目的进展。评述了国内外现代水产种业科技的发展趋势,预言科技创新定会大大促进水产种业的快速发展。

关键词：水产种业发展历程生物技术育种

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跨越新世纪的海洋生物高技术前沿

高技术通讯 2000年第7期10卷 1-4页

作者：相建海中国科学院海洋研究所青岛266071

结合我国实际 ,就新世纪海洋生物高技术前沿和应用领域进行了阐述。

关键词：海洋生物学生物工程分子学

Virus in male reproductive system affect spermiogenesis in marine shrimps Penaeus vannamei and Metapenaeus affinis

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Virus in male reproductive system affect spermiogenesis in m...

中国明对虾凝血栓蛋白基因全长cDNA的克隆及其表达特征

中国海洋与湖沼学会甲壳动物学分会、中国动物学会、中国海洋与湖沼学会生态学分会2000年学术研讨会

作者： Wu Changgong xiang jianhai Institute of Oceanology, Chinese Academy of Sciences Qingdao 266071

Virus in male reproductive system of cultured marine shrimpPenaeus vannamei and wild marine shrimp,Metapenaeus affinis *** can be found in testis and vas *** ***,the infection of virus was serious,there were not maturedsperms were found in the lumen of vas deferens in matured male *** months after sampling,all the cultured shrimps died ***

关键词： Virus Penaeus vannamei Metapenaeus affinis male reproductive system

来源： cnki会议评论

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高技术通讯 2006年第9期16卷 964-969页

作者：王兵李富花谢玉素相建海中国科学院海洋研究所青岛266071 中国科学院研究生院北京100039

获得了中国明对虾凝血栓蛋白（TSP）基因全长cDNA,该基因由2868个碱基组成,具有一个2763bp的开放阅读框,编码920个氨基酸,其中包含21个氨基酸组成的信号肽.中国明对虾TSP由四类不同的结构域组成,从N端开始顺序为几丁质结合结构域,EGF-l... 详细信息

获得了中国明对虾凝血栓蛋白（TSP）基因全长cDNA,该基因由2868个碱基组成,具有一个2763bp的开放阅读框,编码920个氨基酸,其中包含21个氨基酸组成的信号肽.中国明对虾TSP由四类不同的结构域组成,从N端开始顺序为几丁质结合结构域,EGF-like结构域,Type Ⅲ repeat和TSP carboxyl-terminal domain（TSP-C结构域）.结构域分析发现,中国明对虾的N端不具有TSP N-terminal domain（TSPN结构域）,但有一个几丁质结合结构域,其C端的Type Ⅲ和TSP-C结构域非常保守.该基因在弧菌感染后的对虾淋巴器官和肝胰脏中的表达量显著增加,并具有不同的时空表达趋势,提示中国明对虾TSP基因在免疫反应中具有重要作用.

关键词：中国明对虾凝血栓蛋白弧菌感染基因表达