Theoretical strategies for high-resolution mapping of complex genetic disorders in humans

Positional cloning of gene(s) underlying a complex disease trait poses requirement of a highresolution linkage map between the disease locus and genetic marker loci. Recent researches have shown that this may be achieved through appropriately modeling and screening linkage disequilibrium between the candidate marker locus and the major trait locus. However, these models were restricted to the circumstances where genotyping at the disease locus was feasible. The major limitations of pedigree-based linkage analyses were addressed in the light of positional cloning and positional candidate gene identification in humans. It summarizes the recent efforts in developing theories for fine-scale mapping of genes underlying complex genetic variations where the one-to-one relationship no longer exists between phenotype of the genetic disorders and the corresponding genotype. Dedicated to Dr. C. C. Tan for his 90th birthday.     

Molecular characterization of diapause hormone inHelicoverpa armigera

Diapause hormone (DH) is a neurohormone which is secreted from suboesophageal ganglion and responsible for induction of embryonic diapause in many insects. Using rapid amplification of the cDNA ends (RACE) method, the cDNA encoding diapause hormone inHelicoverpa armigera, a main kind of crops pest was cloned. The nucleotide sequence reveals that the mRNA encodes an open reading frame and the 25-aa DH peptide is localized at N-terminal region just after the signal peptide. A homology search showed thatH. armigera DH has high homology with the diapause hormone ofBombyx mori andHelicoverpa zea; and it also belongs to the FXPRL neuropeptide family. Thus,H. armigera DH seems to be a new type of diapause hormone molecule.     

微量量热法筛选促菌特效营养药的研究

用热活性检测仪测定了枸杞、黄芪、灵芝、鹿茸对金黄色葡萄球菌促进作用的热功率—时间曲线;计算了不同浓度营养药物作用下的生长速率常数;根据生长速率常数和浓度的数据,确定了营养药物的最佳浓度.     

棒状杆菌表达载体pJL23的构建

质粒ｐＧＡ４６－４带有从谷氨酸棒杆菌染色体上分离到的有启动功能的片段,用ＰＣＲ技术从ｐＧＡ４６－４中扩增了该片段的关键区域;启动子ＰＧＬ,将该启动子经ＥｃｏＲⅠ－ＢａｍＨ Ⅰ双酶切后,与大肠杆菌质粒ｐＪＬ０１的ＥｃｏＲⅠ－ＢａｍＨⅠ大片段连接,再接入Ｘｙｌ Ｅ ｇｅｎｅ和棒状杆菌质粒ｐＸＺ１０１４２,构建成棒状杆菌－大肠杆菌穿梭表达载体ｐＪＬ２３。用邻苯二酚双加氧酶基因检测表明,该表达载体可在棒状     

Cloning the Promoter of BcNA1 from Brassica napus and Fad2 Gene from Arabidopsis thaliana and Construction of the Plant Expression Vector

The upstream regulatory region of a seed-specific gene was isolated from the genomic DNA of Brassica napus by PCR amplification. The cloned fragment contained 1755 nucleotides, and shared a sequence homology of 99.6% with the reported data. The coding region of oleic acid desaturase gene was then cloned from Arabidopsis thaliana. The sequencing analysis indicated that the sequence of the PCR product was just the same as reported before. In addition, the plant expression vector harboring the seed-specific promoter and trans-Fad2 gene was constructed.     

皮肤渗透促进剂氮酮的合成研究

以己内酰胺为原料,采用相转移催化合成了皮肤渗透促进剂氮酮。应用正交实验技术,研究了原料配比、催化剂用量及时间对反应收率的影响,在优化条件下收率可达８９．９％。     

助剂对自热转化催化剂Ni/γ-Al2O3性能的影响

利用常压固定床微反色谱联用装置及程序升温还原的方法 ,考察了添加不同助剂对自热转化催化剂的还原性能、活性和水热稳定性的影响。实验结果表明 ,稀土氧化物作为助剂添加到模型催化剂中 ,可以大大改善催化剂的初活性和水热稳定性 ,MnO ,CeO2 和La2 O3 是提高催化剂水热稳定性的较好助剂 ,同时CuO是改善催化剂还原性能的一种很好的助剂     

动物克隆研究进展

动物克隆随着"多莉"羊的诞生而逐渐受到人们的关注.动物克隆的核心是细胞核的再程序化.从动物克隆的技术流程、理论基础以及应用领域等方面作一综合论述.     

基于遗传算法的启动子序列多粒度结构分析

采用遗传算法对启动子序列数据集抽取不同长度的最优模糊子序列模式,以此分析启动子的结构。实验发现：适当调整子序列模式与样本序列的匹配程度阈值,可以发现局部频繁模式,除较长的TATA盒外还发现了其他一些模式,而在阈值较低时算法得到CpG岛。     

兔脑组织因子基因在大肠杆菌中的克隆与表达

重组组织因子(recombinant tissue factor,rTF)是新一代凝血酶原时间检测(PT)试剂的主要成分.从新鲜兔脑组织提取总RNA,通过RT-PCR得到TF全长编码基因,并克隆到pMD19-T载体.以测序验证正确的重组子为模板,扩增TF基因并构建成融合表达质粒pMAL-c2X-TF,并将其转化到DH5α中.SDS-PAGE和Western blotting证明,菌体诱导后形成可溶性的融合表达产物,该融合蛋白通过亲和层析纯化和凝血因子X a水解成为游离的TF蛋白,该蛋白通过脂化,制备成重组PT试剂.凝血试验表明,该试剂对血浆有明显的促凝活性.