引物退火控制技术在差异表达基因克隆中的应用

许多年以来,分离差异表达基因的方法仅限于差异筛选cDNA文库,直到1992年Liang等[1]发明了一种检测基因转录模式的方法,即mRNA差异显示技     

海带雌性原生质体的分离与培养

于1993年1月 - 1993年4月，为了制备遗传性纯一的基因工程受体，用酶法从海带雌配子体单性生殖系分离雌性单倍体原生质体。从皱纹盘鲍消化腺及消化器官提取出鲍酶，在25°C下、1％（W／V）的鲍酶与2％（W／V）的纤维素酶（Onozuka R－10）合用能有效解离单细胞和原生质体。海带雌性原生质体经2d培养再生了细胞壁。     

对虾一种无包涵体杆状病毒病原的PCR检测

为建立对虾无包涵体杆状病毒的快速诊断技术,应用PCR技术分别对暴发性流行病发生前、流行中和发病后的对虾样品进行了检测.根据已克隆的对虾杆状病毒部分基因组序列而设计的引物能特异性地扩增出靶DNA片段,最低可以检测1pg的病素DNA.由典型发病症状对虾的胃、腮、肝胰腺、中肠、游泳足、肌内和心脏等器官和组织中成功地检测到病毒.不同组织和器官经扩增得到的信号强弱不同:病虾的胃扩增得到的信号最强,中肠、肌肉、心脏和游泳足次之,腮和肝胰腺信号最弱,说明病毒在不同组织和器官中数量及感染程度不同.在对虾发病前及发病后,用二次PCR还能检测表面不发病呈隐性感染状态的对虾携带的病毒;而对野生健康虾的检测结果为阴性.研究表明,PCR是检测对虾暴发性流行病快速、灵敏而准确的方法,对病毒病的早期诊断、防治和高健康对虾品种的选育具有指导意义.     

Computation of Viscous Uniform and Shear Flow over A Circular Cylinder by A Finite Element Method

The incompressible viscous uniform and shear flow past a circular cylinder is studied. The two-dimensional Navier-Stokes equations are solved by a finite element method. The governing equations are discretized by a weighted residual method in space. The stable three-step scheme is applied to the momentum equations in the time integration. The numerical model is firstly applied to the computation of the lid-driven cavity flow for its validation. The computed results agree well with the measured data and other numerical results. Then, it is used to simulate the viscous uniform and shear flow over a circular cylinder for Reynolds numbers from lO0 to lO00. The transient time interval before the vortex shedding occurs is shortened considerably by introduction of artificial perturbation. The computed Strouhal number, drag and lift coefficients agree well with the experimental data. The computation shows that the finite element model can be successfully applied to the viscous flow problem.     

海面的电磁散射模型及应用

本文采用随机格林函数方法求解了电磁场随机波动方程,给出了具有倾斜表面含气泡海浪的体散射系数表达式,并且指出,当电磁波近垂直入射时,只应用面散射模型是不充分的,必须考虑体散射效应.同时讨论了当入射角在20°～30°之间体散射系数与面散射系数的加权对接问题,给出了完整的海面散射系数计算过程.     

辽河盆地大民屯凹陷流体压力特征

大民屯凹陷是辽河断陷内4个下第三系凹陷之一。在综合利用钻井、试井及地震等资料的基础上，系统研究并论述了大民屯凹陷流体压力特征。基于57口井的声波测井资料，凹陷内泥岩压力特征可区分为正常压力、异常压力或强超压等类型；根据152口井391个点的压力测试数据，凹陷内产油层段的压力梯度多接近于1；利用公式法模拟计算了47条地震剖面的流体压力、剩余压力及压力系数的分布特征，凹陷内剖面压力系统自上而下一般由正常压力、弱超压和强超压3部分组成。此外，还根据流体压力演化的基本原理及钻井、岩性与试井等实际资料，模拟恢复了大民屯凹陷的压力演化史，其可划分为超压原始积累、超压部分释放及超压再积聚3个阶段。总体上，大民屯凹陷的超压强度低于渤海湾盆地其他地区的超压强度。     

天然气水合物的热流特征及影响因素

热流特征是研究天然气水合物的重要指标之一，不同地质环境下的天然气水合物的热流特征不尽相同。天然气水合物的热流分布主要受地壳年龄和沉积作用、地形及断裂构造和流体迁移等因素影响。     

钝顶螺旋藻部分原生质体及单细胞的制备与培养

于1992年1月-1994年4月，进行超声波处理方法制备钝顶螺旋藻部分原生质体以作为基因工程受体，以及制备单细胞用于固体平板克隆化培养的研究。研究结果表明：超声波以20kHz频率、15W功率作用30s，可使藻丝体断裂成15．0±1．6个细胞长度；延长作用时间，至2－6个细胞长度时，细胞壁结构遭到破坏，形成部分原生质体；继续作用，可形成少量原生质体和大量单细胞。断裂藻丝体、部分原生质体、单细胞以及原生质体均可涂布于固体培养基上再生或生长。以一定密度涂布单细胞与原生质体，能够形成彼此分开的单个克隆，可用于筛选及遗传分析。本文提供了一种节省溶菌酶的制备螺旋藻透性体的方法，超声波作用利于外源基因的导入，而涂布培养利于进一步的筛选和形成克隆。     

Dynmnic Response Analysis for Embedded Large-Cylinder Breakwaters Under Wave Excitation

A numerical model is developed for dynamic analysis of large-cylinder breakwaters embedded in soft soil. In the model, the large cylinder is taken as a rigid body divided into elements and the soft soil is replaced by discrete 3D nonlinear spring-dashpot systems. The numerical model is used to simulate the dynamic response of a large-cylinder breakwater to breaking wave excitation. The effects of the dynamic stress-strain relationship models of the soil, the radius and embedded depth of the cylinder, the nonlinear behaviors of the soil, and the limit strength condition of the soil on the dynamic responses of the large-cylinder structure are investigated with an example given. It is indicated that the above-mentioned factors have significant effects on the dynamic responses of an embedded large cylinder breakwater under breaking wave excitation.     

Second-Order Solutions for Random lnterfacial Waves Under Steady Uniform Currents

In the present research, the study of Song (2004) for random interracial waves in two-layer fluid is extended to the case of fluids moving at different steady uniform speeds. The equations describing the random displacements of the density interface and the associated velocity potentials in two-layer fluid are solved to the second order, and the wave-wave interactions of the wave components and the interactions between the waves and currents are described. As expected, the extended solutions include those obtained by Song (2004) as one special case where the steady uniform currents of the two fluids are taken as zero, and the solutions reduce to those derived by Sharma and Dean (1979) for random surface waves if the density of the upper fluid and the current of the lower fluid are both taken as zero.