两种盐对质粒DNA提取的影响

随着基因治疗及基因疫苗的不断发展,质粒的需求量迅速增加。通过以pcDNA3为目的产物,考察了在其提取过程中硫酸铵和氯化钙的加入对pcDNA3收率的影响和对杂质的去除效果。实验发现:硫酸铵和氯化钙的加入都可以明显降低质粒提取液中蛋白质、RNA杂质的含量;但是氯化钙在降低蛋白质和RNA含量的同时也降低了质粒DNA的收率。硫酸铵是去除蛋白质、RNA杂质的良好试剂,同时能够保持较高的质粒DNA收率。当质粒提取液中硫酸铵浓度加入到2.5M时,质粒的纯化因子可达到10.2,而收率在90%以上。     

重组人脂联素基因构建、表达、纯化及活性鉴定

根据脂联素cDNA序列设计并合成9条寡聚核苷酸片段,经重叠PCR技术获得重组人脂联素基因.构建pET-22b( )/ADPN表达载体,转化大肠杆菌BL21(DE3),IPTG诱导表达.目的蛋白以包涵体的形式表达,表达量占菌体总蛋白的30%以上.表达产物通过Ni-NTA柱纯化,SDS-PAGE分析显示,其分子量约为30 ku,与预期结果一致.内皮细胞生长抑制实验证实,重组人脂联素生理浓度下具有抑制人内皮细胞生长的活性并呈时间依赖性.以上工作为进一步研究脂联素的生物学功能奠定了基础.     

Glu101对保持中心蛋白正常构象作用的研究——光谱法研究稀土离子与八肋游仆虫中心蛋白作用对蛋白构象的影响

通过位点直接突变法将八肋游仆虫中心蛋白101位保守的Glu突变成Lys,得到突变蛋白E101K.利用疏水性探针TN S研究了突变对Tb^3＋与蛋白作用时蛋白构象变化的影响.结果表明,相对于钙饱和的蛋白,铽的饱和对蛋白构象变化的影响更大.同时,共振光散射研究表明,Glu101对保持蛋白适当的构象变化行为起着至关重要的作用.     

水稻16kDa醇溶蛋白启动子克隆及载体构建

水稻种子16 kDa醇溶蛋白是水稻种子成熟过程中,由16 kDa基因编码,16 kDa醇溶蛋白启动子调控,在胚乳中特异表达的蛋白质.以水稻基因组DNA为模板,通过PCR扩增技术得到16 kDa启动子片段,序列分析结果表明:获得的启动子片段的大小为931 bp,与已报道的该启动子序列相比较,其核苷酸序列同源性为99.9%.该启动子区域含有TATA-box,CAAT-box,GCN4基序,Prolamin-box等胚乳特异表达启动子所必需的正调控元件.利用该启动子构建了植物种子特异表达载体pC16 kDP.     

G蛋白偶联受体研究进展

G蛋白偶联受体(Gprotein-coupled receptors,GPCRs)是具有7个跨膜螺旋的蛋白质受体,是人体内最大的蛋白质家族。按GPCRs一级结构的同源性,主要分为A、B、C三族;按氨基酸序列的相似性以及与配基的结合情况GPCRs可以分为5个亚家族。本文就GPCRs的结构、分类、GPCRs的二聚化以及其固有活性等方面做了一些介绍。     

海洋生物黏附蛋白研究进展

海洋附着生物粘合蛋白的研究对于生物粘合剂的开发及海洋防污行业均具有重要的意义。综述了对贻贝足丝蛋白,藤壶胶蛋白以及管栖毛虫管胶蛋白的研究进展。3种不同生物的黏附蛋白具有某些相似性,同时又各有其结构特点,对其蛋白质结构,功能及粘合机制的研究既具有重要的学术意义也具有广阔的应用前景。     

植物细胞膜H^＋-ATPase对必需矿质养分的适应性

植物细胞膜H^＋-ATPase是初级转运蛋白，在各种离子和代谢产物的跨膜运输中起着重要的作用；介绍了植物细胞H^＋-ATPase的结构、生化特性、生理功能与调控机制；重点综述了细胞膜H^＋-ATPase的活性及其蛋白数量与基因表达对植物必需矿质养分的适应性。     

Scanning for sisnatures of geosraphically restricted selection based on population genomics analysis

Natural selection, as the driving force of human evolution, has direct impact on population differentiation. However, it is still unclear to what extent the genetic differentiation has been caused by natural selection. To explore this question, we performed a genome-wide scan with single nucleotide polymorphism （SNP） data from the International HapMap Project. Single locus FsTanalysis was applied to assess the frequency difference among populations in autosomes. Based on the empirical distribution of FsT, we identified 12669 SNPs correlating to population differentiation and 1853 candidate genes subjected to geographic restricted natural selection. Further interpretation of gene ontogeny revealed 121 categories of biological process with the enrichments of candidate genes. Our results suggest that natural selection may play an important role in human population differentiation. In addition, our analysis provides new clues as well as research methods for our understanding of population differentiation and natural selection.     

Comparing the biological coherence of network clusters identified by different detection algorithms

Protein-protein interaction networks serve to carry out basic molecular activity in the cell. Detecting the modular structures from the protein-protein interaction network is important for understanding the organization, function and dynamics of a biological system. In order to identify functional neighbor- hoods based on network topology, many network cluster identification algorithms have been devel- oped. However, each algorithm might dissect a network from a different aspect and may provide dif- ferent insight on the network partition. In order to objectively evaluate the performance of four com- monly used cluster detection algorithms: molecular complex detection (MCODE), NetworkBlast, shortest-distance clustering (SDC) and Girvan-Newman (G-N) algorithm, we compared the biological coherence of the network clusters found by these algorithms through a uniform evaluation framework. Each algorithm was utilized to find network clusters in two different protein-protein interaction net- works with various parameters. Comparison of the resulting network clusters indicates that clusters found by MCODE and SDC are of higher biological coherence than those by NetworkBlast and G-N algorithm.