青岛文昌鱼肌球蛋白重链基因片段的重组表达

将青岛文昌鱼(Branchiostoma belcheri tsingtauense)肌球蛋白重链基因片段亚克隆到pET-30a质粒,构建出重组表达载体并转化入大肠杆菌Escherichia coli BL21中.经SDS-PAGE和Western杂交检测表明,在IPTG诱导下含有重组载体的菌株可表达分子质量约30 ku的融合蛋白.诱导条件优化实验结果显示,该菌株经0.2 mmol/L IPTG诱导1 h就可大量表达此蛋白.可溶性实验确认该重组蛋白是可溶性蛋白.本研究为文昌鱼肌球蛋白重链特异性抗体的制备奠定了基础.     

自组织竞争神经网络在江苏油田有杆抽油系统故障诊断中的应用

有杆抽油系统的故障诊断技术是国内外采油工程技术中的一个重要研究课题。通过示功图的不同形状特征可以反映抽油机的不同工作状态。将自组织竞争神经网络应用于示功图的识别与分类,建立了一个自组织竞争神经网络模型对示功图进行自动聚类,从而实现故障诊断的自动化。应用江苏油田的实测示功图数据进行实验,可以看出自组织竞争神经网络具有良好的分类能力和泛化性能,是实现油田抽油系统故障诊断的有效技术,具有很强的实用价值和广泛的应用前景。     

用显微注射方法分析牙鲆肌肉发育调节基因MyoD, Myf5的启动子序列

克隆了牙鲆(Paralichthys olivaceus)肌肉发育调节基因MyoD和Myf5的启动子序列,分别为608 bp和1 073 bp。对序列进行分析发现MyoD和Myf5的启动子含有参与肌肉发育调控的基因的结合位点。将MyoD和Myf5的启动子连接到GFP载体上,构建了重组质粒MyoDP-GFP和Myf5P-GFP,并注射到一细胞或两细胞的斑马鱼胚胎中,对这两个基因的启动子进行了瞬时表达研究。结果表明,牙鲆MyoD和Myf5的启动子可以驱动绿色荧光蛋白特异地在斑马鱼肌肉纤维中表达,因此牙鲆608 bpMyoD和1073 bpMyf5的启动子包含着可以驱动MyoD和Myf5正常表达的必需元件,它们是肌肉特异的,并且可以跨物种行使其功能。     

Quantitative comparison of the expression of myogenic regulatory factors in flounder (Paralichthys olivaceus) embryos and adult tissues

MyoD, Myf5, and myogenin are myogenic regulatory factors that play important roles during myogenesis. It is thought that MyoD and Myf5 are required for myogenic determination, while myogenin is important for terminal differentiation and lineage maintenance. To better understand the function of myogenic regulatory factors in muscle development of flounder, an important economic fish in Asia, real-time quantitative RT-PCR was used to characterize the expression patterns of MyoD, Myf5, and myogenin at early stages of embryo development, and in different tissues of the adult flounder. The results show that, Myf5 is the first gene to be expressed during the early stages of flounder development, followed by MyoD and myogenin. The expressions of Myf5, yoD, and myogenin at the early stages have a common characteristic: expression gradually increased to a peak level, and then gradually decreased to an extremely low level. In the adult flounder, the expression of the three genes in muscle is much higher than that in other tissues, indicating that they are important for muscle growth and maintenance of grown fish. During embryonic stages, the expression level of MyoD might serve an important role in the balance between muscle cell differentiation and proliferation. When the MyoD expression is over 30% of its highest level, the muscle cells enter the differentiation stage.     

Quantitative comparison of the expression of myogenic regulatory factors in flounder (Paralichthys olivaceus)embryos and adult tissues

MyoD,Myf5,and myogenin are myogenic regulatory factors that play important roles during myogenesis.It is thought that MyoD and Myf5 are required for myogenic determination,while myogenin is important for terminal differentiation and lineage maintenance.To better understand the function of myogenic regulatory factors in muscle development of flounder,an important economic fish in Asia,real-time quantitative RT-PCR was used to characterize the expression patterns of MyoD,Myf5, and myogenin at early stages of embryo development,and in different tissues of the adult flounder.The results show that,Myf5 is the first gene to be expressed during the early stages of flounder development, followed by MyoD and myogenin.The expressions of Myf5,MyoD,and myogenin at the early stages have a common characteristic:expression gradually increased to a peak level,and then gradually decreased to an extremely low level.In the adult flounder,the expression of the three genes in muscle is much higher than that in other tissues,indicating that they are important for muscle growth and maintenance of grown fish.During embryonic stages,the expression level of MyoD might serve an important role in the balance between muscle cell differentiation and proliferation.When the MyoD expression is over 30%of its highest level,the muscle cells enter the differentiation stage.     

牙鲆Myogenin蛋白在大肠杆菌中重组表达的研究

采用大肠杆菌表达系统及pProEXTMHTc表达载体对牙鲆肌肉发生调节因子myogenin基因进行了体外重组表达研究。通过对诱导时间、IPTG诱导浓度以及诱导温度的优化,确定了最佳诱导条件为:诱导温度31℃,IPTG终浓度0.6mmol/L,诱导时间2 h。在该条件下,重组蛋白表达量最高且是可溶性的。Myogenin融合蛋白的成功表达为进一步深入研究其在牙鲆肌肉发育过程中的分子机制奠定了基础。     

基于GIS的河南省县级城市空间聚类研究与实现

将GIS与空间聚类算法相结合,可以从空间数据集中发现对象的凝聚趋势、分布规律和发展方向,并可进一步挖掘分析,从而获取更加概括和精练的信息。本文以河南省108个县级城市为研究对象,对其三年的人口及从业人员等状况进行聚类分析。采用系统聚类中的离差平方和法,以欧式距离的平方作为度量标准,对变量以Z得分作为转换标准,最终把样本划分为5类,并制作聚类划分结果专题地图。结合聚类分析和聚类分布图,进一步证实了空间聚类在县级城市空间分析中的可用性,能为河南县级城市的发展规划制定提供重要依据。