对虾白斑综合症病毒极早期基因启动子筛选文库的构建

对虾白斑综合症病毒（white spot syndrome virus,WSSV）是危害对虾养殖业的主要病原,而WSSV极早期基因对病毒功能基因的转录翻译和调控起着至关重要的作用.本研究将WSSV基因组以超声法随机断裂至400～900bp DNA片段,并插入启动子缺失且多克隆位点下游具有报告基因的载体来构建文库,转染昆虫细胞Hi5.倒置荧光显微镜观察发现部分转染后的细胞有增强型绿色荧光蛋白（EGFP）的表达,表明这部分细胞中的DNA片段可能具有极早期启动子活性.流式细胞仪分析显示,这部分荧光细胞占总细胞的比例为0.35%.由于转染效率为50%,可认为占比0.7%的WSSV基因组随机片段能在Hi5细胞中激发下游报告基因的转录翻译,可能具有极早期基因启动子活性.这对于进一步筛选和鉴定WSSV极早期基因及开展极早期启动子研究具有重要的意义.     

Preparation by enzymolysis and bioactivity of iron complex of fish protein hydrolysate （Fe-FPH）from low value fish

Preparation of Fe^2＋ chelate of fish protein hydrolysate （Fe-FPH） obtained from low value fish proteins was introduced and its bioactivity was studied by compound enzymolysis. The optimum conditions for hydrolysate chelating Fe^2＋ are DH （degree of hydrolysis） at 5%, pH 7.0, 20℃ and 15 min chelating time for FM （material not being defatted）. Four types of Fe-FPH including CA （deposit after chelating）, CB （deposit in 50% of absolute ethanol solution）, CC （suspended deposit in 80% of absolute ethanol solution）, and CD （bottom deposit in 80% of absolute ethanol solution） were fractionated with absolute ethanol from FM. Structural analysis through infra-red spectrum revealed that Fe^2＋ was combined strongly with amino-group and carboxyl-group in each chelate and each Fe^2＋ could form two five-member ring structures. All of the four chelates were shown more significant antioxidative activity and can be used as natural hydrophobic and hydrophilic antioxidant. Among all the chelates, the CB possesses the most effective antioxidative activity at 92% as high as that of a-tocopherol. Among all Fe-FPHs, only CD showed the most effective antibacterial activity against Escherichia coli, Staphylococcus aureus, Salmonella typhi, and Bacillus subtilis and can be used as natural antibacterial. It provides a more effective way for utilization of low value fish proteins and key information of Fe-FPH as additive in food industry.     

凡纳滨对虾G蛋白Gαq基因的克隆和功能鉴定

为了寻找并克隆凡纳滨对虾(Litopenaeus vannamei)G蛋白α亚基,为研究对虾的生理调控提供理论基础,通过简并PCR和RACE反应获得目的基因的全长cDNA序列;用Blast, DNAstar和Genedoc软件对所得序列进行分析,确定所得序列为凡纳滨对虾G蛋白Gαq基因,命名为pvGαq.将该基因的编码序列克隆到表达载体,通过免疫共沉淀实验和胞内Ca2 , IP3浓度的测定鉴定该Gαq亚基的功能,发现该亚基的序列和功能与其他Gαq家族成员具有高度保守性.用Western blotting分析发现pvGαq在对虾身体各部位存在普遍分布,尤其在脑神经、眼柄、腮和颚片中有大量表达,在嗅觉器官触角也有适量分布.说明了pvGαq在对虾生命活动中的重要性,为研究对虾的生理调控奠定了理论基础.     

湖泊沉积物中蛋白质和氨基酸的动态变化

测定了阿哈湖和百花湖沉积物孔隙水中蛋白质和氨基酸的含量。蛋白质含量分别在0．32－1．71mg/ml和0．22－1．38mg/ml之间，氨基酸含量分别在1．15－5．01μg/ml（以含N量计）和1．41－4．16μg/ml之间。两湖沉积物氨基酸含量逐渐增高，表明扩散作用的存在。随着时间的推移，蛋白质在沉积物各层中得到了很好的保存，说明它的分解比较有限。沉积物孔隙水中氨基酸含量和微生物活动有关，氨基酸在27cm以后的阿哈湖沉积物中明显积累，可能指示微生物活动的减弱。     

甘肃窑街中侏罗世化石角质层有机质及其碳同位素组成与古环境的关系

通过对甘肃窑街中侏罗统窑街组化石银杏以及现代银杏角质层有机质组分和碳同位素组成分析．表明银杏角质层有较强的抗风化能力．化石和现代银杏角质层有机质烷烃和甾萜类化合物组成十分相似。可以通过角质层的碳同位素组成等参数进行古环境重建，结果表明中侏罗世早期古环境较为温湿．到了中期后古气候逐渐变得干旱少雨，后期降雨量又逐渐增加，温度升高，植被发育。     

Divergence of Parartemia and Artemia haemoglobin genes

Parartemia is a genus of brine shrimp endemic to Australia which is related to the more widespread and economically importantArtemia sp. The expression of a multimeric haemoglobin molecule inArtemia is well documented but inParartemia only trace levels of a possible haemoglobin have been observed. In this paper we describe the DNA sequence of a domain of a haemoglobin molecule inParartemia. The derived amino acid sequence suggests that the possible date of divergence about 85 million years ago of the two genera predates the divergence of the C and T polymers ofArtemia haemoglobin. This date would correlate with the physical and temporal isolation of Australia in the late Mesozoic.     

Characterization of the particulate organic matter in the Loire Estuary (France) using ETS activity measurements

The major problems encountered in studying estuarine particulate organic matter (POM) are the characterization and quantitation of different kinds of materials (e.g. detrital and living matter, algal and bacterial matter…) each type contributing a specific role in the ecosystem. The study of the activity of the electron transport system (ETS) is proposed as a tool for resolving these problems. Results obtained in our laboratory with cultures of planktonic algae and bacteria provided us with numerical relationships between ETS activity and various organic components such as chlorophyll, carbon and protein. These relationships were: ETS activity/chlorophyll = 2.6, carbon/ETS activity = 17.3, protein/ETS activity = 9.6 in algae; carbon/ETS activity = 5.9, protein/ETS activity = 7.7 in bacteria (ETS activity expressed in /i₂h⁻¹ at 20°C; chlorophyll, carbon and protein in μg). Such data can be applied in field studies to characterize the living algal and bacterial matter and, by difference, the detrital organic matter. We report here a study of the Loire estuary as an example.     

东方百合(Lilium)杂交品种索邦病程相关蛋白(PR1)基因的克隆与原核表达

病程相关蛋白（PR）广泛参与植物对生物和非生物胁迫的应答。采用cDNA末端快速克隆技术,从东方百合（Lilium）杂交品种索邦中克隆了PR1基因,并命名为LhSorPR1。LhSorPR1基因cDNA全长870 bp,5’非翻译区和3’非翻译区的长度分别为47 bp和274 bp。该基因的开放阅读框为549 bp,编码182个氨基酸。成熟的LhSorPR1蛋白（不含信号肽）分子量为18.19 kDa,等电点为5.69。采用同源建模方式构建的LhSorPR1蛋白三维结构与其模板番茄PR1蛋白p14a具有较高的相似性,说明百合PR1蛋白在结构和功能上与其他植物中的PR1蛋白具有较强的保守性。构建pCold Ⅱ-LhSorPR1原核表达,经IPTG诱导后,目的蛋白在大肠杆菌中以包涵体形式成功表达。使用免疫印迹方法对表达的LhSorPR1重组蛋白进行了验证,并对带有His标签的重组蛋白成功进行了纯化。