一种用磷酸钙法高效转染HEK 293T细胞方法的建立

利用磷酸钙转染法将lacZ-pShuttle质粒导入HEK 293T细胞中,实验证明,pH值、DNA纯度、转染后培养时间和甘油休克时间是影响转染效率的重要因素.转染后培养6h进行甘油休克效果最佳,比培养0h直接进行休克转染效率升高584.4%;甘油休克时间为4.5min时转染效果最佳,比不用甘油休克转染效率升高130.8%.通过对影响转染的几个条件进行优化,建立了1种可以高效转染HEK 293T细胞的方法,且简便易行.     

Rapid detection of virulent protease secreted by Vibrio anguillarum by dot enzyme-linked immunosorbent assay

Dot enzyme-linked immunosorbent assay （dot-ELISA）, indirect ELISA and Westem blot were performed to detect the virulent protease secreted by Vibrio anguillarum which was isolated from the diseased left-eyed flounder, Paralichthys olivaceous. Sensitivity results showed that dot-ELISA is a more sensitive, rapid and simple technique for the protease detection. The minimal detectable amount of protease is about 7 pg in the dot-ELISA test, while 7.8 ng in the indirect ELISA and 6.25 ng in the Westem blot respectively. Protease could be detected 2 h after incubation of V. anguillarum in the 2216E liquid medium but enzyme activity was very low at that period. From 6 to 12 h, the amount and enzyme activity of protease increased markedly and reached maximum at stationary phase. Analysis of serum samples periodically collected from the infected flounders showed that after 2 h of infection by V. anguillarum, the pathogenic bacteria could be detected in the blood of the infected flounders but no protease was found. It was 5-6 h after infection that the protease was detected in blood and then the amount increased as infection advanced. Quantitative detection of protease either incubation in the medium or from the blood of infected flounders could be accomplished in virtue of positive controls of quantificational protease standards （＂marker＂） so that the alterations ofprotease secretion both in vitro and in vivo could be understood generally. In addition, the indirect ELISA and dot-ELISA were also performed to detect V. anguillarum cells. Results indicated that the sensitivity of indirect ELISA to bacteria cells is higher than that of the dot-ELISA, and that the minimal detectable amount is approximately 10^4 cell/mL in the indirect ELISA, while 10^5 cell/mL in the dot-ELISA.     

江苏岸外辐射沙脊群东沙稳定性研究

东沙是江苏岸外辐射沙脊群中的第二大沙洲,具有独特的地形地貌和水动力条件,对它进行稳定性研究为揭示整个辐射沙洲及其邻近岸滩的动态演变都非常有益。通过利用多年遥感卫片资料、1998年取得的现场水文泥沙观测资料和东沙滩面表层沉积物资料等,对东沙的地形地貌特征、沉积特征和东沙两侧潮汐通道的水流泥沙特征等进行了详细分析。研究结果表明,东沙的沙脊偏于西侧,西侧滩面较窄、高程较高且岸线较为顺直,东侧滩面较宽、高程较低且岸线较为破碎;西洋和陈家坞槽均处于冲刷状态,净输沙的主要方向为输向槽外或输向条子泥;东沙近三十年来面积有所缩小且有外围向中央收缩的趋势,尤其以向东、向南方向的迁移最为明显。     

东海原甲藻cDNA文库构建及尝试性EST分析

东海原甲藻(Prorocentrumdonghaiense)是中国沿海频繁发生的大规模赤潮的原因种之一,大规模分离鉴定东海原甲藻功能基因是理解东海原甲藻赤潮形成过程和机理的重要基础。取对数生长期藻体,经微量总RNA抽提、cDNA合成、cDNA扩增和克隆等步骤,构建了东海原甲藻cDNA文库并进行了尝试性表达序列标签分析。从含有约5000个转化子的文库中随机取150个测序,获得126条EST序列。经网上BlastN及BlastX分析,共发现11个是已知功能的基因的标签。这些基因与东海原甲藻生长发育、物质转换和能量代谢相关。     

金属离子对锯缘青蟹NAGase活力的影响

本文研究了几种金属离子对锯缘青蟹（Scylla serrata）N-乙酰-β-D-氨基葡萄糖苷酶（NAGase）活力的影响．其结果表明：Li^＋、Na^＋和K^＋对酶活力没有明显影响，Mg^2＋、Ca^2＋和Ba^2＋对该酶均有激活作用，激活程度依次为Ca^2＋〉Ba^2＋〉Mg^2＋．Al^3＋、Fe^3＋、Cd^2＋、Pb^2＋和Hg^2＋对该酶具有一定的抑制作用，2．0μmol／dm^3的Hg^2＋可以使酶活力完全丧失．Co^2＋对酶的效应是先激活后抑制，Mn^2＋对酶仅有轻微的激活作用．Cd^2＋和Fe^2＋对锯缘青蟹NAGase的抑制作用都呈竞争性-反竞争性混合Ⅱ型效应，Cd^2＋的抑制常数Ki和KiS分别为23．9、5．0mmool／dm^3，Fe^3＋的抑制常数Ki和KiS分剐为395．5、135．6μmol／dm^3．Ki〉KIS说明酶一底物络合物（ES）与抑制剂的亲和力比游离酶（E）与抑制剂的亲和力大．     

Studies on Genus Cryptomonas from China seas

ＩＮＴＲＯＤＵＣＴＩＯＮＣｒｙｐｔｏｍｏｎａｓｉｓｕｎｉｃｅｌｌｕｌａｒｂｉｆｌａｇｅｌｌａｔｅｗｈｉｃｈｉｓａｃｏｍｍｏｎｇｅｎｕｓｏｆＣｒｙｐｔｏｐｈｙｔａ .Ｔｈｅｃｅｌｌｓａｒｅｕｓｕａｌｌｙｓｔｒｏｎｇｌｙｃｏｍｐｒｅｓｓｅｄｌａｔｅｒａｌｌｙ .ＴｈｅｒｅａｒｅｏｎｅｏｒｔｗｏｌａｒｇｅＣｈｌｏｒｏｐｌａｓｔｓｔｈａｔｃｏｎｔａｉｎｃｈｌｏｒｏｐｈｙｌｌａ ,ｃ ,ｓｏｍｅｃｈａｒａｃｔｅｒｉｓｔｉｃｃａｒｏｔｅｎｏｉｄｓａｎｄｐｈｙｃｏｂｉｌｉｎｐｉｇｍｅｎｔｓ (Ｈｕｅｔａｌ.,1 997;Ｗｅｉ…     

碳水化合物的组合合成

碳水化合物的组合合成是一项新兴技术，该技术可以在短时间内合成大量用于进行生物活性筛选的寡糖及拟糖物。液相与固相合成技术可以极大地加快药物研究与开发进程。作者针对该技术在碳水化合物合成方面的研究进展情况进行讨论。     

天然热释光技术在海洋油气田勘查中的应用

沉积物热释光能有效地反映海洋环境下深部油气田引起的微弱放射性异常。热释光在不同的测试粒级、不同的岩性中的丰度不同，结果为粗粒级＞细粒级、砂岩＞粉砂质砂岩＞泥质粉砂岩＞泥岩。热释光也与部分金属元素含量有关，Ti、Ba、Zn、Fe^3 /Fe^2 与热释光呈正相关关系。热释光与烃类也呈较强的正相关性。不同采用深度热释光测量结果表明，热释光剂量随沉积物的沉积深度和沉积时间的增加而增加，深层样的热释光对油气藏的指示效果更好。     

夜光藻的室内培养

夜光藻的室内培养对于研究夜光藻的生理生态特性及其增殖机理是一个关键性的环节。通过试验，发现在实验室条件下以扁藻作为饵料，夜光藻能良好的生长和繁殖，从而为夜光藻的室内培养解决了关键性的饵料问题，实现了国内夜光藻室内的长期培养。     

PREDICTION OF MOTION OF SEMI-SUBMERSIBLE DRILLING PLATFORM IN MAVES

Starting from potential theory, this paper presents a new procedure for computing the hydrodynamic forces and six-degrees-of-freedom motion for a semi-submersible platform at an arbitrary heading in waves by using strip theory and linearized viscous damping hypothesis, taking the effects of the free surface and the interaction between main hulls into account. The differential equations of motion for the floating platform are formulated in a new way and the associated computer programs are developed. Comparisons between the theoretical computing results and the full-scale and model-scale experiments show that the present method gives more accurate results than those by previous methods.