Cloning, Expression of Histone 2A from Scylla paramamosain and Analyses of Antimicrobial Activity and Mechanism of synthetic peptide Sphistin

拟穴青蟹（Scyllaparamamosain）是我国南部沿海重要经济养殖品种，由于缺乏拟穴青蟹的相关免疫知识，迄今无法真正实现拟穴青蟹的健康养殖。抗菌肽类的免疫因子在海洋动物的先天性免疫中具有重要作用，因此挖掘新的抗菌肽对揭示拟穴青蟹的免疫机制具有重要的科学意义。组蛋白Histone是真核生物染色体的基本结构蛋白，一些研究证明组蛋白的成分具有抗菌活性。本实验室在前期研究中，从LPS刺激的拟穴青蟹血细胞SSHcDNA文库中筛选到一个类似Histone2A的基因，推测其可能在拟穴青蟹体内具有抗菌等免疫功能，因而对其进行了细致的研究工作，取得了以下研究成果。 1.阐明了拟穴青蟹Histone2A...Mud crab (Scylla paramamosain) is an important economic aquaculture variety of China's southern coast. It is still difficult to artificial culture of S.paramamosain because its immune knowledge is unclear. The antibacterial peptide plays an important role of congenital immune in Marine animal’s immune factors. Finding new type of antibacterial peptide has important scientific significance to the i...学位：理学硕士院系专业：海洋与环境学院环境科学与工程系_环境科学学号：2262008115152

Relative Quantification of Hepcidin Gene Transcripts in Pseudosciaena crocea

以大黄鱼(PSEudOSCIAEnA CrOCEA)管家基因18S rrnA和β-ACTIn作为内参基因,分别比较2个内参基因建立的相对定量曲线,最终确立以18S rrnA为参比基因,定量分析大黄鱼的HEPCIdIn抗菌肽基因.该相对定量分析方法所得结果与nOrTHErn-blOT方法一致.应用建立的HEPCIdIn基因的实时荧光相对定量研究方法,对大黄鱼头肾中的HEPCIdIn基因转录物进行相对定量,为今后开展鱼体内免疫相关基因的表达特性、诱导机制等工作奠定研究基础.同时,克隆得到的大黄鱼β-ACTIn基因和18S rrnA基因片段已提交基因库,并获得登录号.Two house-keeping genes,18S rRNA and β-actin,were cloned as endogenous genes for relative quantification of an antimicrobial peptide hepcidin in large yellow croakers(Pseudosciaena crocea) using the real-time RT-PCR.In comparision of the slopes of the relative standard curves between the two endogenous genes and the target gene,a relative quantification method for hepcidin gene using 18S rRNA as the calibrator gene was established.Results obtained with this method were consistent with the rusults from Northern-blot,suggesting feasibility and effectiveness of the method.This paper provides an effective method to investigate expression patterns and induction profiles of the hepcidin gene in large yellow croakers,and other immunity-related genes in fish bodies.The cloned fragments of 18S rRNA and β-actin genes from the large yellow croaker have been submitted to the GenBank,which have provided accession numbers.福建省科技计划重点资助项目(2007I0022);上海市高校选拔优秀青年教师科研专项基金资助项目(SHU08021);上海大学创新基金资助项目(A10011109006

The Effects of Benzo [a] Pyrene (BaP) Exposure on the CYP1A1 mRNA and AhR2 mRNA Expression of Red Seabream(Pagrus major)

通过水体暴露方式对海水养殖真鲷进行bAP持续染毒,利用实时定量PCr技术研究了真鲷细胞色素P450基因(CyP1A1)和芳香烃受体基因(AHr2)随bAP暴露剂量、时间的动力学变化。结果发现,0.1~1.5μg/l环境浓度的bAP能够显著性诱导CyP1A1基因和AHr2基因的表达,且AHr2 MrnA早于CyP1A1 MrnA被诱导表达;bAP持续暴露48 H,CyP1A1和AHr2基因的表达水平均随暴露时间的延长而显著升高,染毒72 H后又回复到本底水平,实验表明这两个基因的表达与bAP的暴露剂量和暴露时间之间具有显著性的剂量-效应和时间-效应关系。The gene expression patterns of CYP1A1 and Aryl hydrocarbon receptor(AhR2) of red seabream(Pagrus major) were both measured using real-time quantitative PCR(qPCR) when fish exposed to environmentally relevant concentration of BaP(0.1,0.5 and 1.0 μg/L,respectively).The results showed that CYP1A1 mRNA and AhR2 mRNA could be induced significantly,besides,the time of AhR2 mRNA induced ahead of the time of CYP1A mRNA induced.The two genes were induced markedly at the begining of BaP exposure,and then decreased to the basal levels after 72 h.The results demonstrate that BaP can regulate CYP1A1 and AhR2 transcript in a dose and time dependent manner.国家“863”计划重点资助项目(2007AA091406);国家自然科学基金资助项目(30770391

Benzo[a]pyrene modulation of acute immunologic responses in red Sea bream pretreated with lipopolysaccharide

The effects of polycyclic aromatic hydrocarbons (PAHs) have been reported to modulate the immune response in aquatic animals, but the collected information of their effects on fish immunity is so far ambiguous. This study demonstrated that Benzo[a]pyrene (BaP) exposure altered the expression pattern of an antimicrobial peptide hepcidin (PM-hepc) gene and the activities of some immune-associated parameters in the lipopolysaccharide (LPS)-challenged red sea bream (Pagrus major). It was observed that LPS could increase respiratory burst, lysozyme and antibacterial activity in P. major. However when the P. major was exposed to different concentrations of BaP (1, 4, or 8 μg L-1) for 14 days and then challenged with LPS there was no significant change in the lysozyme and antibacterial activity. It was further observed that LPS could induce the PM-hepc mRNA expression at 3, 6, and 12-h post-LPS challenge. However, when P. major was exposed first to BaP for 14 days and then challenged with LPS, the expression of PM-hepc mRNA was delayed in the liver until 24 h and not significantly induced until 48 and 96 h. The mRNA expression pattern was completely different from that only with LPS challenge, showing that BaP exposure changed the PM-hepc mRNA expression pattern of fish with LPS challenge. This study demonstrated that BaP exposure can weaken or inhibit the induction of lysozyme and antibacterial activity in the LPS-challenged P. major; conversely BaP exposure could enhance the mRNA expression of PM-hepc gene, indicating that the effect of BaP has different modulatory mechanism on hepcidin genes and immune-associated parameters. ? 2012 Wiley Periodicals, Inc

透射法测量丰中子核~8He的去中子截面

利用HIRFL提供的50MeV/u~ (13)C束流轰击Be靶,通过RIBLL选择出丰中子放射性核~8He．利用透射法测量了~8He的去双中子截面和去四中子截面．利用Ogawa等人的理论结合原有~8He的反应总截面数据,得到~8He的核芯为~4He的结论

中能区丰中子核~(11)Li的反应总截面的测量(英文)

利用HIRFL提供的50MeV/u的~(13)C束流轰击Be靶,通过RIBLL选择出放射性核素~(11)Li.实验采用透射法测量了25—45MeV/u的~(11)Li在~(28)Si靶上的反应总截面.采用双参数Gauss密度分布形式,利用Glauber模型很好地拟合了高能和中能区的~(11)Li实验数据,并从密度分布中提取了核的物质均方根半径