A novel methodology for finding the regulation on gene expression data

DNA microarray technology is a high throughput and parallel technique for genomic investigation due to its advantages of simultaneously surveying features of large scales complex data in biology. This paper aims to find feature subset to build the classifier for gene expression data analysis. At first, K-means clustering algorithm was carried out on the dataset of yeast cell cycle. Based on rand calculation, a statistical method was used to pick out the data points (genes) for classifier design. Meanwhile, the principal component analysis was applied to help to construct the classifier. For the validation of classifier built and prediction of a target subset of genes, discriminant analysis in terms of partial least square regression and artificial neural network were also performed.     

牙鲆fibrinogen β基因的克隆及表达分析

通过mRNA差异显示发现一个鳗弧菌刺激后在牙鲆肝脏中表达量显著增加的片段,结合RACE技术得到了该差异片段1 856 bp的全长cDNA,包含一个1 479 bp的开放阅读框,编码的蛋白质含有fibrinogen β的C末端签名序列.同源性分析表明其氨基酸序列与鱼类以及哺乳类的fibrinogen β都具有高度的相似性,因此可断定此基因为牙鲆的fibrinogen β基因(GenBank登录号为EF581895).RT-PCR分析表明,在注射鳗弧菌后,该基因在肝脏、肾脏、脾脏、腮、肠、心脏中的表达量呈现逐渐增加的趋势.推测其可能在鳗弧菌侵染中,起到了促进伤口愈合的作用,或通过和其他细胞因子结合在防御细菌的侵染中发挥免疫调节作用.     

IL-21/4-IgG4融合基因的真核表达

利用新型细胞因子IL-21的变异体IL-21/4与IgG4构成融合基因IL-21/4-IgG4. 将构建好的融合基因载体poptivec- IL-21/4-IgG4转染到中华仓鼠卵巢细胞中,成功并稳定表达了IL-21/4-IgG4融合蛋白.     

绿色木霉EG Ⅲ基因的克隆及其在工业酿酒酵母中的整合表达

通过RT-PCR从绿色木霉AS3.3711中克隆得到EG Ⅲ基因,序列分析显示该基因编码序列全长为1 257 bp,编码418个aa,且自身带有21个aa的信号肽序列,与里氏木霉eg3的同源性为99.6%.将该基因连入酿酒酵母多拷贝整合型表达载体pScIKP中获得重组质粒,线性化后电转化酿酒酵母工业菌株AS2.489,通过SDS-PAGE蛋白电泳、刚果红染色和酶活测定对转化子进行了分析.结果表明:转化子有明显的重组EG Ⅲ表达带,能够产生CMC水解圈,说明eg3基因已在酿酒酵母中获得了正确的分泌表达,表达EG Ⅲ的重组菌产酶高峰出现时间为60 h,最高酶活接近120 U/mL,重组酶EG Ⅲ的最适温度为60 ℃,最适pH为6.0,转化子的遗传稳定性达到99.17%,实现了绿色木霉eg3基因在工业酿酒酵母中的稳定高效表达.     

绿色木霉纤维二糖水解酶Ⅱ基因的克隆及其在酿酒酵母中的高效表达

采用RT-PCR 的方法从目前最高效的纤维素酶产生菌之一的绿色木霉AS3.3711中克隆得到纤维二糖水解酶Ⅱ（CBHⅡ）的编码基因cbh2。序列测定和分析表明,该编码序列长1416 bp,编码472个氨基酸残基,与已公布的绿色木霉CICC 13038的cbh2仅在第263位存在一个氨基酸残基的差别（Gln替换为Pro）,而与里氏木霉VTT-D-80133的cbh2则完全同源。该序列已经提交GenBank,登录号为 DQ864992。利用信号肽预测软件SignalP 3.0发现该基因自身带有信号肽序列,其最大可能的切割位点在18和19位氨基酸残基之间。将该基因片段接入酿酒酵母高拷贝数整合型表达载体pScIKP中,得到重组表达质粒pScIKP-cbh2。经酶切线性化后电转化酿酒酵母AS2.489菌株,通过G418浓度梯度筛选出高抗转化子。转化子经蛋白电泳分析,发现该基因的信号肽序列能够被酿酒酵母识别,因此重组CBHⅡ成功分泌到了胞外。但可能由于酿酒酵母表达系统的过度糖基化作用,重组CBHⅡ比出发菌绿色木霉的CBHⅡ分子量要高许多,并且可能因为过度糖基化作用位点不同而形成了不同大小分子量的表达产物。转化子经CMC糖化力法测定酶活,发现该重组CBHⅡ酶活性并没有受过度糖基化作用的影响,最高可达7.71 U/mL,比大多数报道的要高。其作用的最适pH值为5.0,最适温度为65 ℃,且具有较好的热稳定性。     

基于非负矩阵分解的面部表情识别研究

提出了一种基于非负矩阵分解与支持向量机相结合的面部表情识别方法。使用直方图均衡化等方法对人脸图像进行预处理,使用非负矩阵分解算法进行表情特征提取,采用支持向量机对面部表情进行分类。以Matlab为仿真工具,在日本女性人脸表情数据库上测试。取得了66.19％的识别率。     

唐宋词体功能的文化迁延

娱乐、抒怀与社交是词的主要功能。晚唐、五代和北宋词主要是以音乐形式应用于娱乐场合。北宋中期以后,随着词在社会上的广泛流行,其社交功能逐渐增强,宴词、寿词大多是社交活动的产物。进入南宋,国事变迁,词的创作与传播环境发生变化,抒怀言志成为词的主要功能之一。     

论《诗经》中动物物象的情感表达作用

《诗经》中的动物物象在先民的社会情感交流、政治化的情义沟通和幽默化的人生呈现中,扮演着重要角色。各种各样的动物物象传递着先民的爱情、亲情和友情,也传递着政治关系中的信任和建议,更是先民幽默人生态度的最佳代言。从诗歌的角度,则是加强了艺术表现的力度,丰富了艺术表现的手法。     

Effects of Chinese herbs capable of replenishing qi, nourishing yin and activating blood circulation and their compatibility on differentially expressed genes of ischemic myocardium

This study was conducted to investigate the effects of Chinese herbs capable of replenishing qi, nourishing yin and activating blood circulation and their compatibility on differentially expressed genes of ischemic myocardium which were selected from differential expression profile we had established before, and to explore the underlying mechanism. The acute myocardial infarction （AMi） model was established by ligating the left anterior descending （LAD） coronary artery, then the model rats were randomly divided into the model group, the Metoprolol group, the replenishing qi nourishing yin （RN） group, the activating blood circulation （AB） group, and the replenishing qi, nourishing yin and activating blood circulation （RA） group. In addition, the normal group and the sham group were set up. The rats of medication groups were administered by intragastric gavage with corresponding drugs on the second day after operations, and the rats of the normal group and the sham group were given normal saline as the same time.Then the ischemic hearts were harvested on the 8th day after operation. The myocardial pathomorphological changes were observed under a light microscope. The mRNA changes of target genes such as COX5a and ATP5e were detected using Real-time fluorescence quantitative PCR （Q-PCR）, and the activities of related enzymes were detected by colorimetric assay. The main resuits were as follows： the histological changes were observed by HE staining, and cardiocyte swelling, inflammatory cell infiltration and cytolysis were showed in regional ischemic myocardium of the model group, while the pathomorphological changes in all medication groups did not show obvious changes. Two genes related to energy metabolism, COX5a and ATP5e, were selected as the target genes which were down-regulated at the mRNA level in the medication groups. The activities of correlative functional enzymes also decreased in the RA group compared to that in the model group accordingly （P〈0.05）. The results indicated that the abnormal expression of genes involved in energy metabolism pathways could be one of the molecular mechanisms of AMI. The compatibility of Chinese herbs capable of replenishing qi, nourishing yin and activating blood circulation affects the expression of energy-relative gene COX5a, ATP5e, which is probably the mechanism of myocardial preservation, and is more effective than single herb of replenishing qi and nourishing yin or activating blood circulation.