蓝杆藻ATCC51142中琥珀酸半醛脱氢酶的原核表达及生化表征

【背景】蓝藻中生成琥珀酸的三羧酸循环途径与其他物种不同。由于α-酮戊二酸脱羧酶和琥珀酸半醛脱氢酶的存在使得蓝藻的三羧酸循环途径变得完整。琥珀酸半醛脱氢酶催化琥珀酸半醛氧化为琥珀酸,在蓝藻中广泛存在。【目的】克隆、表达和纯化蓝杆藻ATCC51142中cce4228基因编码蛋白,并对其进行生化表征。【方法】以蓝杆藻ATCC51142基因组为模板克隆得到cce4228基因,将其插入到原核表达载体pET-28a上,在大肠杆菌BL21(DE3)细胞中进行异源表达,利用Ni-NTA树脂纯化cce4228蛋白。运用紫外分光光度法和生物信息学方法表征重组cce4228蛋白生化特性。【结果】构建了pET-28a-cce4228重组表达质粒,重组cce4228蛋白在大肠杆菌中得到可溶性表达,获得了纯度大于90%的cce4228蛋白。酶动力学测试和生物信息学分析结果显示,cce4228蛋白是一个NADP+-依赖型的琥珀酸半醛脱氢酶。【结论】蓝杆藻ATCC51142中cce4228基因编码一个偏好NADP+辅因子的琥珀酸半醛脱氢酶,cce4228蛋白的生化表征结果为进一步深入研究cce4228蛋白的结构功能关系及催化机制奠定了基础。     

Expression changes in the stroma of prostate cancer predict subsequent relapse

Biomarkers are needed to address overtreatment that occurs for the majority of prostate cancer patients that would not die of the disease but receive radical treatment. A possible barrier to biomarker discovery may be the polyclonal/multifocal nature of prostate tumors as well as cell-type heterogeneity between patient samples. Tumor-adjacent stroma (tumor microenvironment) is less affected by genetic alteration and might therefore yield more consistent biomarkers in response to tumor aggressiveness. To this end we compared Affymetrix gene expression profiles in stroma near tumor and identified a set of 115 probe sets for which the expression levels were significantly correlated with time-to-relapse. We also compared patients that chemically relapsed shortly after prostatectomy (<1 year), and patients that did not relapse in the first four years after prostatectomy. We identified 131 differentially expressed microarray probe sets between these two categories. 19 probe sets (15 genes overlapped between the two gene lists with p<0.0001). We developed a PAM-based classifier by training on samples containing stroma near tumor: 9 rapid relapse patient samples and 9 indolent patient samples. We then tested the classifier on 47 different samples, containing 90% or more stroma. The classifier predicted the risk status of patients with an average accuracy of 87%. This is the first general tumor microenvironment-based prognostic classifier. These results indicate that the prostate cancer microenvironment exhibits reproducible changes useful for predicting outcomes for patients.     

大壁虎与嗅觉相关的学习记忆

壁虎科动物鼻腔较大,化学感觉系统发达,类似于对化学信息敏感的舌端游离的蜥蜴目动物（Malan,1946),并且嗅黏膜面积很大,感觉器神经元多,嗅神经系统十分完善（Gabe and Saint,1976).     

苹毛丽金龟对植物挥发物成分的触角电位及行为反应

为筛选适宜的苹毛丽金龟Proagopertha lucidula (Faldermann)引诱剂化学物质, 对已报道的金龟子科两性引诱剂单一组分进行了触角电位（EAG）和Y型嗅觉仪中的嗅觉反应测定。结果表明: 在10 μg刺激剂量下测试的11 种植物挥发物中, 只有顺-3-己烯-1-醇、香叶醇、茴香脑和丁香酚有明确的EAG反应值, 其他7种化合物具有弱反应值。剂量反应曲线表明, 具有明确的EAG活性的4种化合物激发的反应强度与化合物的剂量呈正相关。在Y型嗅觉仪中, 顺-3-己烯-1-醇、香叶醇、茴香脑和丁香酚4种化合物在1, 10, 100 μg剂量下的行为反应百分率都显著高于对照溶剂正己烷, 且雌雄之间无显著差异。该研究结果可为开发苹毛丽金龟田间引诱剂奠定基础。     

日本血吸虫未成熟卯单链抗体库的构建、筛选及初步应用

运用噬菌体展示技术构建日本血吸虫未成熟虫卵可溶性抗原（SIEA）单链抗体（scFv）表达文库,以天然分子候选疫苗SIEA26～28ku为靶抗原筛选SIEA单链抗体库,获得特异性单链抗体．并将该scFv基因亚克隆至原核高效表达载体PET32a,诱导SIEA26-28ku特异性scFv大量表达．随后以此为探针筛选日本血吸虫尾蚴cDNA文库,以期获得SIEA2628ku天然分子候选疫苗相关的编码基因．结果显示,所获得的SIEA26-28ku特异性scFv,表达量高,采用该探针初步筛选出相关基因核糖体蛋白S4．SIEA26-28ku特异性scFv的获得,为进一步筛选、分析鉴定抗日本血吸虫病天然分子候选疫苗SIEA26-28ku的编码基因奠定了基础．     

Structural properties of replication origins in yeast DNA sequences

Sequence-dependent DNA flexibility is an important structural property originating from the DNA 3D structure. In this paper, we investigate the DNA flexibility of the budding yeast (S. Cerevisiae) replication origins on a genome-wide scale using flexibility parameters from two different models, the trinucleotide and the tetranucleotide models. Based on analyzing average flexibility profiles of 270 replication origins, we find that yeast replication origins are significantly rigid compared with their surrounding genomic regions. To further understand the highly distinctive property of replication origins, we compare the flexibility patterns between yeast replication origins and promoters, and find that they both contain significantly rigid DNAs. Our results suggest that DNA flexibility is an important factor that helps proteins recognize and bind the target sites in order to initiate DNA replication. Inspired by the role of the rigid region in promoters, we speculate that the rigid replication origins may facilitate binding of proteins, including the origin recognition complex (ORC), Cdc6, Cdt1 and the MCM2-7 complex.     

Ectopic expression of <Emphasis Type="Italic">OsMADS1</Emphasis> caused dwarfism and spikelet alteration in rice

In rice, an E-class gene, OsMADS1, acts to specify the identities of the lemma and palea. In this study, the OsMADS1 gene with a CaMV35S promoter was transformed into a japonica cultivar, Zhonghua 11. All transgenic plants successfully showed similar phenotypes, including dwarfism, distorted panicles, decreased numbers of branches and spikelets, and elongated sterile lemma. Histological analysis showed that the elongated sterile lemma developed with silicified epidermal and sclerenchymal cells, which were lacking in the wild-type sterile lemma, suggesting that the elongated sterile lemma had assumed the identity of the lemma or palea. Some marker genes were subjected to a detailed analysis of the distribution of their expression among the lemma, palea and sterile lemma. DROOPING LEAF (DL) and OsMADS6 genes were only expressed in the normal lemma or palea, respectively. In the elongated sterile lemma, a high level of DL gene expression was detected, while no expression of OsMADS6 was found, implying that the sterile lemma transformed into the lemma but not the palea. These results provide clues to elucidate the mechanism of evolution from lemma to sterile lemma in rice. qPCR analysis also suggested that the ectopic expression of OsMADS1 induced abnormal brassinosteroid and gibberellin acid activation, and then resulted in developmental defects in the stem and panicle.     

中亚热带典型人工林常见乔灌木根际效应——以江西泰和千烟洲为例

为了深入探讨人工林内不同植物根际效应的差异, 为人工林生态系统林下植被管理提供理论依据, 该研究以江西泰和千烟洲站区杉木(Cunninghamia lanceolata)、马尾松(Pinus massoniana)和湿地松(Pinus elliottii) 3种人工林(1985年前后营造)为研究对象, 测定林内乔木及其林下常见灌木檵木(Loropetalum chinense)、杨桐(Adinandra millettii)和格药柃(Eurya muricata)的根际和全土土壤碳、氮、磷含量及pH值, 比较不同林分类型下优势乔灌树种的根际效应。结果发现: (1)乔木根际土壤大部分化学性质指标显著高于或低于全土(p < 0.05), 而林下灌木根际土壤与全土土壤化学性质的差异与物种有关, 如檵木根际土壤大部分化学性质显著不同于全土, 而格药柃根际土壤大部分指标与全土无显著差异。(2)除硝态氮(NO₃ ^--N)外, 林下灌木不同物种之间的根际效应有别, 具体表现为檵木pH值、铵态氮(NH₄ ⁺-N)、可溶性有机碳(DOC)、全氮(TN)、全碳(TC)、碳氮比(C/N)、有效磷(AP)和全磷(TP)的根际效应显著高于格药柃, TN、TC、C/N和AP的根际效应也显著高于杨桐, 但杨桐与格药柃间的根际效应无显著差异。(3)杉木的根际效应显著强于林下3种灌木; 马尾松和湿地松的根际效应与檵木无显著差异, 而马尾松的根际效应显著高于杨桐和格药柃, 湿地松的根际效应显著高于格药柃。该研究表明相对于林下灌木(尤其是格药柃), 乔木具有较强的根际效应, 暗示了乔木具有更高的养分捕获能力。但林下灌木与乔木根际效应的差异与灌木种类和林分类型有关。因此, 人工林林下植被管理除了考虑适量保留林下灌木外, 可依据人工林类型, 充分考虑灌木种类选择, 进而最大地发挥人工林生态系统的生产和生态功能。     

蛋白质结构型的定义和识别

提出紧结构域的概念,由二级结构序列中一段或几段连续的α螺旋和β折叠构成的空间紧密堆集的最大折叠体称为紧结构域.利用3种紧结构域（α域,β域和α/β域）定义球蛋白的5种结构型：α型蛋白,β型蛋白,α/β型蛋白,多域蛋白和ζ型蛋白.将1 261个代表性的蛋白质(1 022家族)进行分类,并和SCOP库的分类做了比较.进行了删去序列冗余的分析.在此基础上提出结构型的预测方案,成功率在82%～85%.