Cloning and identification of Na+/H+ antiporter gene(nhaA) from Pseudomonas sp. & Oryza sativa transformation

Na+/H+逆向转运蛋白（Na+/H+antiporter）NhaA(以下简称NhaA)具有逆浓度梯度向胞外或液泡转运Na+的特性，从而使该生物或细胞能在较高的盐度下生存。根据原核基因无内含子的特点，本实验参照大肠杆菌的nhaA基因的序列设计引物，以极端嗜盐细菌假单胞杆菌（Pseudomonassp.cn4902）的总DNA为模板，通过PCR扩增出一新结构基因，并将它克隆至pMD18-T载体上。测序结果表明：该结构基因长1089bp，编码362个氨基酸。通过生物信息学方法，经GenBank中Blast比较得知，本序列与多种生物的nhaA基因高度同源，与E.coliK12的nhaA基因的同源性高...Na+/H+ antiporters are membrane proteins that play a major role in pH and Na+ homeostasis of cells throughout the biological kingdom, either pump Na+ out of the cells actively or translocate it from cytoplasm into vacuoles in the plant cells. This is one of the key reasons that bear responsibility for the organisms surviving in a high salinity environment. According to the essential character that...学位：理学硕士院系专业：生命科学学院生物学系_生物化学与分子生物学学号：20012603

Effect of ZnF2 Coating on Performance of LiNi0.5Mn1.5O4 Cathode Material for Lithium-ion Batteries

采用溶胶-凝胶法制备了LiNi0.5Mn1.5O4正极材料，并利用Zn F2对其表面进行包覆改性。XRD、SEM和TEM测试表明，包覆处理不影响材料的晶体结构，2%(质量分数，以LiNi0.5Mn1.5O4质量计，下同)的Zn F2在LiNi0.5Mn1.5O4表面形成了约7 nm厚均匀包覆层。对未包覆的LiNi0.5Mn1.5O4和1%、2%、3%的Zn F2包覆后的LiNi0.5Mn1.5O4的电化学性能进行了考察，发现Zn F2包覆能够减弱电解液与LiNi0.5Mn1.5O4正极材料之间的相互作用，稳定电极表面，提高材料的电化学性能。其中，2%Zn F2包覆样品表现出最佳的循环性能和倍率性能，0.2C电流倍率下循环200圈后，其放电比容量维持在109.0 m A·h/g，保持率为79.7%；5 C电流倍率下循环500圈后，放电比容量维持在94.2 m A·h/g，保持率为85.6%

Advances in the Catalysts and Mechanism for Ammonia Synthesis

通讯联系人：廖代伟[中文摘要]本文综述了氨合成这一最重要的工业多相催化过程８０ 多年来的研究进展。介绍了氨合成熔铁催化剂、氨合成钌催化剂的发展以及对氨合成催化反应机理的不同看法。[英文摘要]The advances in ammonia synthesis as is one of the most important industrial heterogeneous catalytic processes are reviewed. The developments of both fused iron catalysts and ruthenium catalysts and the different opinions on catalytic mechanism for ammonia synthesis are discussed.国家自然科学基金;固体表面物理化学国家重点实验室和福建炼油厂资

Raise of the Frequency in Green-shoot-formation for Rice Anther Cultivation

福建省水稻攻关课题基金 (K790 81);; 厦门市科委科技发展项目 ( 3 5 0 2 2 2 0 0 0 10 4

粉末电致发光的新应用──一种光神经器件

近年来，人工神经网格与神经网络计算机的研究在世界范围内已形成一个热点，本文通过对电致发光材料的各种特性的讨论，分析了固体化平板电致发光作为一种光神经器件的一些特点，指出了它在神经网络实现中有很好的应用前景

Cloning and Characterization of Na~+/H~+ Antiporter Gene (nhaA) from Pseudomonas sp.cn4902

根据3种生物的Na+/H+逆向转运蛋白基因(nhaA)的两端序列设计引物,利用PCR从假单胞菌 (Pseudomonassp.cn4902)中克隆得到一结构基因。该基因长1089bp,编码362个氨基酸,与E.coliK12的 nhaA基因的同源性高达97.0%。将该结构基因与pBV220构建成重组载体pBVA。SDS PAGE电泳表明:含 pBVA的转化子产生较高浓度的分子量约为41kD的蛋白,与预期相符。在含NaCl1.0mol/L的培养基中生长达 到平衡期时,转化子的菌浓度约是对照的2.3倍。经原子吸收光谱测定,转化子细胞质中Na+浓度仅为对照菌的 60.4%。SDS PAGE电泳表明该基因的表达蛋白位于细胞膜(壁)上。提纯外源基因表达蛋白并对其N端8个氨 基酸进行测序,与nhaA基因推测的氨基酸序列完全相符。这些实验证实,克隆得到的基因是假单胞菌的nhaA基 因。该基因已经在GenBank登记,收录号为AY643494。 【英文摘要】 According to the sequences of the gene nhaA coding for Na~+/H~+ antiporter,a structural gene was cloned from Pseudomonas sp.cn4902 by PCR reaction with a set of primers.It was 1 089 bp in length and codes for 362 amino acids sharing homology with the gene nhaA of E.coli K12 as high as 97.0%.It was inserted into plasmid pBV220 to form a high level expression reconstruction plasmid pBVA.So an overexpression 41 kD protein band could be found in the lane of transformant harbored with pBVA after SDS-PAGE electro...福建省科技计划重点资助项目(编号:2003N053);; 厦门大学生命科学学院细胞生物学与肿瘤细胞工程教育部重点实验室项目(编号:2004106)~

CLONING AND EXPRESSION OF MANNITOL-1-PHOSPATE DEHYDROGENASE GENE OF PSEUDOMONAS sp. cn 4902

参照几种生物的甘露醇 1 磷酸脱氢酶基因 (mtlD)的序列设计引物 ,以极端耐盐的假单胞菌 (Pseudomonassp cn 4 90 2 )的总DNA为模板 ,采用PCR扩增、构建重组高效表达载体以及生物信息学研究等方法 ,进行基因克隆、表达及功能定性等研究。结果表明 ,克隆获得一长为 114 9bp的基因。经蛋白质保守区域研究 ,初步判别该基因为mtlD结构基因。将该基因与pBV2 2 0质粒构建成高效表达原核重组载体pBH。SDS PAGE电泳表明 ,含pBH的转化子产生特异的、分子量约为 4 1kD的蛋白带 ,表达量约占菌体可溶性蛋白 6 7%。转化子的耐盐水平比对照提高了约 1/5。在含 0 9mol/LNaCl的液体培养基中 ,转化子培养 2 4h后其生物量约是对照的 10 2倍 ,甘露醇含量约是对照的 4 1倍。可见 ,假单胞菌的甘露醇 1 磷酸脱氢酶基因是一个重要的耐盐相关基因 ,该基因已在GenBank登记 ,代号为AY112 6 96。According to the essential characteristic of the genome of prokaryotes that no intron can be found in the genes,an attempt was made to find a new gene of mannitol-1-phosphate dehydrogenase ( mtl D) in Pseudomonas sp. cn 4902 by means of PCR amplification. Based on the short conserved sequence of the halo-tolerant-related gene in some microorganisms,a pair of primers were designed for PCR amplification with the template of genomic DNA of Pseudomonas sp. cn 4902. An approximate 1. 1kb DNA fragment of PCR amplification product could be found in the agarose gel electropherogram. The fragment was,then,retrieved,a poly A “tail” added and finally lined together with plasmid pMD18-T overnight to form the reconstructed vectors. The white colonies were picked out,some of them were cultivated and the plasmids were extracted in the transformants and digested by two endonucleases Bam HⅠ and Sal Ⅰ for the detection of the reconstructed vectors after agarose gel electrophoresis. The inserted DNA fragment was sequenced and transferred to the multicloned site of high efficiency expression plasmid pBV220 in E.coli JM101 to constitute a new plasmid named pBH. The tolerance levels against to serious concentrations of NaCl and its protein expression products were researched. It was clear that the cloned gene was 1149bp in length and coded for 383 amino acid residuals. Using the biological informatics in GenBank we found that the nucleotide sequence and deduced amino acid residuals of this gene share more than 90% homology with the mtl D gene of other organisms,including E. coli K12. The result of protein SDS-PAGE electrophoresis of transformants indicated that a 41kD protein coincided with the molecular weight of the transferred mtl D gene,which was 6.7% of the total protein,was produced in this transformants under the condition of 42℃ hot shock. As a result,the highest tolerant level against NaCl of the transformants cultivated in the agar LB medium was raised from 0.9 to 1.1mol/L,which was 22% higher than that of the control. Meanwhile,the biomass of transformants cultivated in the liquid medium containing 0.9mol/L NaCl was about ten-fold that of the control. It was believed that the cloned gene should be a mannitol-1-phosphate dehydrogenase ( mtl D) gene,one of the key genes responsible for osmoregulation in the cells of organisms. The successful cloning and expression of the mtl D gene supplied a good foundation for cultivation of halo-tolerant-related crop plants in the future.科技部转基因植物专项基金资助 ,J0 0 B 0 14号 ;; 厦门市科委科技发展基金资助项目 ,35 0 2 2 2 0 0 0 10 4

Cloning,sequencing and expression of glucitol-6-phosphate dehydrogenase(gutD) gene

本研究根据6 磷酸山梨醇脱氢酶基因(gutD)两端序列设计引物,以假单胞杆菌(PseudonmonasXM)的总DNA为模板通过PCR的方法克隆gutD基因并进行序列分析.将克隆得到的gutD基因与原核表达载体pBV220连接并转化大肠杆菌(Es cherichiacoli)JM101后检测其蛋白表达及菌株生长耐盐性.A glucitol-6-phosphate dehydrogenase (gutD) was cloned viaPCR method using Pseudomonas genomic DNA as template. The gutD gene was sequenced and expressed in pBV220 vector. The transformant was tested for its salty tolerance

大数据时代的工具书资源及知识产权保护

工具书资源于科研或教育或文化至关重要，但由于订购或服务形式变化，已经没有在中国本土&ldquo;收藏&rdquo;或&ldquo;保存&rdquo;，使得我国对这类资源的长期可持续利用受到挑战，怎样长期保存与访问这些工具书资源的问题应运而生。因此，本书聚焦工具书资源发展和使用的情况、长期保存现状与面临的挑战、数据资源的知识产权保护等内容，为我国对该类资源的长期保存与保护提供参考。</p

Mutation and identification of mutant strain of Dunaliella bardawil containing highly in β-carotene

采用紫外辐射诱导巴氏杜氏藻Dunaliellabardawil突变,经初筛和复筛,初步获得一株巴氏杜氏藻的突变株,对两者从细胞形态特征、β-胡萝卜素含量、蛋白质SDS-PAGE电泳以及总DNA的RAPD扩增等方面进行了比较分析。结果表明,在相同培养条件下,两者均为卵圆形,但原种直径仅为突变后藻株直径的60%,颜色差异明显;突变株的β-胡萝卜素含量为原种的6 2～15 5倍;两者遗传相似系数为0 868,与突变株的一般要求相符。鉴于此,确认其为高产β-胡萝卜素的新突变株,命名为Dunaliellabardawilvariant4-5Sun＆Liu。 【英文摘要】 A β-carotene-riched mutant of Dunaliella bardawil by ultraviolet radiation has been identified. The mutant strain was found to be larger in diameter than D. bardawil was. The content of β-carotene of the mutant strain reached as high as 6.20% of the dry matter and was 6-14 times than the wild-type original one. Furthermore, several new bands of protein (polypeptide) of the mutant was identified by the protein SDS-PAGE, meanwhile some were disappeared. The RAPD test revealed that the genetic similarity be...福建省科技计划重点项目资助(2003No53