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.     

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。     

我国陆上大中型气田分布规律与有利勘探区带优选

“八五”、“九五”期间我国天然气探明储量幅度增长,大中型气田分布规律的深化研究密切关系到天然气储量的增长势头。克拉通盆地下层序古隆起控气理论和上层序非构造圈闭控气机理是克拉通大中型气田富集分布的基本规律,前陆盆地是我国富气盆地类型之一,其冲断带下盘大型造圈闭群、前缘斜坡岩性尖灭区带与浅层次生气藏、前缘隆起上的断块圈闭群等是该类盆地有利的天然气富集区带。依据充分的地质资料,结合我国天然气勘探实际,提     

GIS支持下的长江口拦门沙泥沙冲淤定量计算

依据1842～1997年10幅不同年代的长江口海图资料,利用地理信息系统和数字化仪进行处理,建立不同时期的长江口水下数字高程模型,以此作为基础资料,实现了从横剖面、深泓线纵剖面、平面变化等不同角度对长江口拦门沙地区滩槽演变、岸线侵蚀、沙岛形成与变迁等进行研究.通过计算河槽容积,实现了对不同时段泥沙冲淤量的计算.结果表明,155a来拦门沙总的趋势是不断淤积,但不同时期淤积速度大不一样,个别时期甚至会发生一定程度的冲刷,这主要与动力条件的波动有关.1842～1997年,共淤积泥沙38.10亿t,平均每年淤积0.246亿t,约占长江来沙的5%,年均淤厚为1.1cm泥沙淤积部位主要在九段沙、横沙及横沙东滩、崇明东滩三处.发生冲刷的范围较小,仅占总面积的21.4%,主要在北槽,北港上段和南槽局部也有轻微的冲刷发生.     

黄河流域2002年麦收期间一次强降水过程分析

本文通过对陕西南部的佛坪县2002年6月8日——lO日的降水达到了489mm的特大暴雨降水过程的初步分析，对进一步了解大尺度环流背境场下的强降水过程的成因，提高对暴雨天气过程的预报能力是非常有益的；本文收集的降水过程期间的天气图和传真图资料对预报员和海洋工程技术人员有参考价值。     

青岛近海地区SO2和SO42-干沉降速率的季节变化

用小麦岛和团岛观测站提供的青岛近海气象和水温资料,以适于SO2气体和硫酸盐(SO42-)气溶胶的干沉降模型研究了模型中的输送阻尼对两种污染物质干沉降速率的贡献,并计算了2003年青岛近海地区两种污染物质干沉降速率的季节变化。结果表明:对SO2而言,粘性副层传输系数对沉降速率的贡献大于空气动力学传输系数;对于SO42-,空气动力学传输系数对沉降速率的贡献要大于表面传输系数。SO2和SO42-的干沉降速率变化范围分别为0.187~0.868cm/s和0.188~0.532cm/s。两种污染物质干沉降速率的四季变化有相似的规律,即冬季>秋季>春季>夏季。     

黄茅海河口湾现代动力地貌体系和冲淤过程分析

1980—1993年对黄茅海河口湾进行沉积物采样和水流测定及水深测量。根据水动力和地形条件，冲淤分析及Mclaren模型研究河口湾的动力地貌体系、冲淤特征和现代沉积物运移。结果表明：（1）水下地形主要为下泄流或上溯流控制的“深槽－槽沟－浅滩－湾口”的动力地貌体系，反映了河口湾“东进西出”的水流格局；（2）整个河口湾以淤积为主，只有崖门深槽有较明显的优势冲刷特征，并随着崖门深槽向海推移和河口湾“东进西出”水动力作用，黄茅海落潮三角洲相应向西南进积；（3）应用Mclaren模型揭示了黄茅海河口湾现代沉积物运移规律，同样反映了河口湾具有“东进西出”的运移趋势。     

Molecular identification of bloom-forming species Phaeocystis globosa(Prymnesiophyta)and its dispersal based on rDNA ITS sequence analysis

Abstract-The colony-forming Phaeocystis species are causative agents of dense bloom occurrences incoastal waters worldwide. It is difficult to separate them because of the different morphologies associatedwith their colonial stages. In this study we applied molecular approaches to analyze the genetic variationof Phaeocystis globosa and Phaeocystis pouchetii from several geogrtaphic regions, and to assist in tracingthe dispersal of bloom-forming Phaeocystis species in coastal waters of China. The Sequences of the inter-nal transcribed sacers (ITS1 and ITS2) of rDNA and the 5.8S ribosomal RNA gene of Phaeocystisstrains were determined. Sequence comparison shows that P.globosa was the most divergent toP. pouchetii, exhibiting sequence divergence higher than 0.08. However, lower genetic divergences ex-isted between strains of P.globosa. The sequence comparison of the Phaeocystis rDNA ITS clearlyshows that the species isolated from the southeast coast of China is identified as P. globosa rather thanP. cf. po     

Application of the first internal transcribed spacer(ITS-1) of ribosomal DNA as a molecular marker to population analysis in farrer''s scallop Chlamys farreri

Sequence variation of the first internal transcribed spacer of ribosomal DNA （ ITS - 1 ） was examined and its application to the study of genetic variation was explored in four populations of farter＇ s scallop Chlamys farreri. ITS - 1 fragments, with a length of about 300 bp,of 78 individuals collected from Dalian, Qingdao, Yantai in China and Korea respectively were amplified via PCR, cloned and sequenced. Intra-genomic variation was examined by sequencing several clones of single individuals. Alignment and polymorphism analysis detected 44 haplotypes and 50 polymorphic sites which consist of 30 substitutions and 20 indels, indicating a high level of polymorphisms. Sequence analysis also showed a very low level of intra-individual variation. All these features validated the feasibility of application of ITS - 1 fragment to population analysis. Polymorphism analysis showed that the Korea sample has the richest genetic variation, followed by Yantai and Qingdao samples. AMOVA （analysis of molecular variance） showed that the majority （96.26%） of genetic variation was distributed within populations and 3.74% resulted from among populations, but with P 〈 0.05 （ = 0.042）, indicating that the populations in this study have significant divergence. This output was basically concordant with the result arising from RAPD data and different from that from mitochondrial 16S rDNA sequence data. Discussion on this inconsistency was made accordingly.