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Cloning, expression and characterization of human tissue-specific DNA polymerase λ2

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science china(life sciences) 2007年第4期50卷 457-465页

作者： GU Fu1, YOU Chun1, LIU JianPing1, CHEN Ao1,,YU Yao1, WANG Xiang1, WAN DaFang2, GU JianRen2, YUAN HanYing1, LI YuYang1 & Lü Hong1 1 state key laboratory of genetic engineering, Institute of genetics, school of life sciences, fudan university, shanghai 2.0433, china 2 state key laboratory of Oncogenes and Related Genes, shanghai Cancer Institute, shanghai 200032, china Present address: Food Inspection and Supervision Institute of Shanxi Province, No. 222 Changzhi Road, Taiyuan 030012, china 1. state key laboratory of genetic engineering Institute of Genetics School of Life Sciences Fudan University Shanghai 200433 China 2. state key laboratory of Oncogenes and Related Genes Shanghai Cancer Institute Shanghai 200032 China

DNA polymerase (POL) λ plays an important role during DNA repair and DNA nonhomologous recom-bination processes. A novel POL λ variant was cloned from a human liver cDNA library and named POL λ2 (GenBank Accession No. AY302442). POL λ2 has 2206 base pairs in length with an open reading frame of 1452 base pairs encoding a 482-amino-acids protein. Bioinformatics analysis reveals that POL λ2 spans 7.9 kb on human chromosome 10q24 and is composed of 8 exons and 7 introns. It has the specific domain of DNA polymerase X family-POL Xc at the C-terminus and BRCT domain at the N-terminus. POL λ2 was localized predominantly in nucleus in transfected L0-2 cells. It was expressed abundantly in liver and testis, weakly in ovary, and undetectably in other tested human tissues. In comparison with the expression ratio between POL λ and POL λ2 in normal liver tissues and hepato-cellular carcinoma (HCC) adjacent tissues, the ratio was aberrant in 80% of those 15 HCC specimens examined due to the up-regulated expression of POL λ. This abnormality might be involved in hepato-carcinogenesis. The recombinant POL λ2 with His-tag was expressed as a soluble active protein in E. coli BL21 (DE3)CONDON Plus and purified by Ni-NTA resin and then desalted by Superdex-75 chro-matography in an FPLC system. The analysis using isotope α-32P-dCTP incorporation in vitro showed that the purified recombinant POL λ2 exhibited DNA polymerase activity.

关键词： POL λ2 gene cloning expression purification DNA polymerase activity hepatocellular carcinoma

人绒毛膜促性腺激素β亚基单一B-细胞表位(β5、β9或β8)融合蛋白的表达和纯化

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生物工程学报 2004年第1期20卷 49-53页

作者：徐万祥何亚萍何远王丽婷孙志达季朝能顾少华陈金中应康谢毅上海市计划生育科学研究所生殖生物学研究室上海200032 复旦大学生命科学院遗传所遗传工程国家重点实验室上海200433

作为开发新型实用性人绒毛膜促性腺激素 (hCG)疫苗的一种尝试 ,我们已构建若干组合靶抗原三个线性B_细胞表位和外源强T_细胞表位的基因工程hCG嵌合肽。为了检测用这些嵌合肽免疫的动物血清中是否能产生抗各表位的三种抗体 ,本研究选用... 详细信息

作为开发新型实用性人绒毛膜促性腺激素 (hCG)疫苗的一种尝试 ,我们已构建若干组合靶抗原三个线性B_细胞表位和外源强T_细胞表位的基因工程hCG嵌合肽。为了检测用这些嵌合肽免疫的动物血清中是否能产生抗各表位的三种抗体 ,本研究选用能在大肠杆菌中高表达和与生物素亲和性强且特异 (方便通过亲和层析纯化 )的链霉亲和素为载体 ,分别构建了三种含 β_hCG不同单一线性B_细胞表位 (β5 ,β9和 β8)的融合蛋白。在链霉亲和素基因下游多克隆区EcoRⅠ和HindⅢ位点插入各表位编码基因片段 (带TAA终止密码子 )的pTSA_18重组质粒 ,转化BL2 1(DE3)pLysS宿主菌后 ,它们在IPTG诱导下均能以较高水平表达各自目的融合蛋白 ,而且它们的表达产物在Westernblot鉴定中都能被抗各表位特异的多抗或单抗或抗报告表位单抗识别。用改良的制备性PAGE方法可以一步纯化电泳均一性高于 95 %的三个融合蛋白 ,它们的收得率相对 1L培养物约为 5mg。作为化学合成表位肽的替代物 ,β_hCG三个单一B_细胞表位融合蛋白的可获得性将有助于所构建hCG基因工程嵌合肽以及其他hCG疫苗 ,也包括它的DNA疫苗的免疫原性分析。

关键词：人绒毛膜促性腺激素 β亚基单线性表位链霉亲和素融合表达免疫印迹纯化

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葱莲凝集素基因的克隆(英文)

西南师范大学学报（自然科学版） 2002年第3期27卷 433-436页

作者：庞永珍姚剑虹申国安唐克轩谈锋复旦大学生命科学学院遗传学研究所遗传工程国家重点实验室上海200433 西南师范大学生命科学学院重庆400715

利用RACE -PCR技术从葱莲叶片中克隆出葱莲凝集素的全长cDNA .通过比较葱莲同其它石蒜科植物的凝集素基因序列和推测的氨基酸序列 ,发现葱莲凝集素基因编码一具有信号肽的前体蛋白 .葱莲凝集素同其它石蒜科植物凝集素一样为具有

关键词： cDNA 凝集素基因 RACE-PCR 葱莲基因克隆石蔚料氨基酸序列基因编码

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紫杉醇提取纯化技术研究进展

中国生化药物杂志 2004年第1期25卷 59-62页

作者：祝顺琴谈锋唐克轩西南师范大学生命科学学院现代生物医药研究所重庆400715 复旦大学遗传工程国家重点实验室上海200433

综述近年来紫杉醇提取纯化技术中的提取、萃取、柱色谱和高效液相色谱几个步骤的研究进展。

关键词：紫杉醇提取纯化综述

Multi-index Synthesize Evaluation Model Based on Entropy Theory for Dialysis Adequacy

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Multi-index Synthesize Evaluation Model Based on Entropy The...

2009 IEEE International Conference on Intelligent Computing and Intelligent Systems(ICIS2009)

作者： Mei Zhang 1 Yi-sheng Shan 2 3 Nan Wu 1 Yue-ming Hu 1 (1.The ministry of education engineering research center for precision electronic manufacture equipment College of automatic science and engineering South china university of Technology 2.Nephrology department Zhongshan Hospital fudan university.shanghai china.3.Nephrology department First Affiliated Hospital Wenzhou Medical College Wenzhou china

Dialysis adequacy shows validity of dialysis schemes in renal failure patients,which is relative to state of *** accurate evaluation for dialysis adequacy is very *** at the clinic actual situation,an evaluation index system of dialysis adequacy is proposed in this *** view of subjectivity in the determination of weights in the traditional model,entropy theory is used to determine the significance of each index and attribute *** the same time,to aggregate group decision opinions,a consensus function of group decision makers or experts is presented,and then expert weights are ***,a multi-index synthesizing evaluation model of dialysis adequacy is established in this *** illustrative example shows the effectiveness and practicability of the proposed model.

关键词： multi-index synthesize evaluation entropy weight dialysis adequacy

The Balanceabililty of aMM Scheduling Policy for GSS ystems

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The Balanceabililty of aMM Scheduling Policy for GSS ystems

第二十九届中国控制会议

作者： WANG Xingxuan1,ZHENG Da-Zhong2 1 department of Electronic engineering,fudan university,shanghai 2.0433,P.R.china2 department of Automation,Tsinghua university,Beijing 100084,P.R.china

The generalized switched server（GSS）system and some switching scheduling policies,such as the FRR,FMM,aMM,and FSS policy,have been proposed for resolving a cluster of task balancing problems,which exist in a wide varietyof engineering *** paper mainly studies the balanceability of the aMM switching scheduling policy for the generalized switched server *** maximum load status difference（MLD）,called as the system difference,is *** is shown that the GSS system would have a smaller MLD under the aMM policy than under the FRR policy,and further,for a given GSS system,the aMM policy is balanceable if the FRR policy is.A general case where the number of the parallel units N is arbitrary is investigated,and a sufficient condition for the balanceability of the aMM policy in the case where the N equals to three is obtained.

关键词： Generalized Switched Server Systems(GSS) Switching Scheduling Policy Balanceability Load Balancing Hybrid Dynamic Systems(HDS) Algebra Constraint

Constructing cDNA Microarray to Isolate Novel Differentially Expressed Genes Related to Human Glioma and Characterizations of Novel Full-length Gene PPIL-3

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Constructing cDNA Microarray to Isolate Novel Differentially...

中国抗癌协会神经肿瘤专业委员会第八届学术会议

作者： Zhen-Yu Qi1,Wang zhong,Yao Li2,Zong-Xiang Zhou2,Guo-Zhen Hui1,Yi Xie2 1.department of Neurosurgery,The First Affiliated Hospital of Suzhou university,Suzhou,215006,People''s Republic of china 2.state key laboratory of genetic engineering,Institute of genetics,fudan university,220 Han Dan Rd.,shanghai,2.0433,P.R.china

Identification of the genes that are differentially expressed between brain tumor and normal brain tissues is important for understanding the molecular basis of these nerve system tumors and for defining possible targets for therapeutic *** objective of my research is to investigate the differentially expressed genes related to human glioma using cDNA microarray and study the characterization of one novel full-length *** RNA was extracted from human glioma tissues and normal brain tissues,and mRNA was used to make *** hybridization and washing procedure,the results of hybridization were scanned using computer *** gene named 681F05 clone was one differentially expressed gene to human glioma through four times hybridizations and *** 681F05 clone was performed by northern blot,in situ hybridization,5’RACE,chromosome mapping and *** differentially expressed genes to human glioma were through four times hybridizations and *** demonstrated that 681F05 clone was low-expression in human brain tissue and over-expression in human glioma *** analysis of BLASTn and BLASTx showed that 681F05 clone is two cDNA clones encoding two novel proteins that are higher identity to the cyclophilin isoform 10 of ***,*** analysis revealed the two cDNA clones are two different splicing variants of a novel cycophilin-like gene（PPIL3a and PPIL3b）.According to the Nomenclature Committee of Human Genome Organization（HUGO）,the gene was named as peptidylprolyl isomerase（cyclophilin）-like 3（approved symbol,PPIL3）.Two nucleotide sequences found in this study have been submitted to the GenBankTM with accession numbers:AF251049 and ***,PPIL3 was located in chromosome 2q33 between STS marker stSG2762（proximal） and SHGC-3074（distal）.We experienced that cDNA microarray technology can be successfully applied to identify differentially expressed genes in human *** approach

关键词： Differentially Expressed Gene Glioma Microarray Novel Full-length Gene

System Control Design for Generalized Switched Server Systems

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System Control Design for Generalized Switched Server System...

第二十九届中国控制会议

作者： WANG Xingxuan1,ZHENG Da-Zhong2 1.department of Electronic engineering,fudan university,shanghai 2.0433,P.R.china 2.department of Automation,Tsinghua university,Beijing 100084,P.R.china

The generalized switched server（GSS）systems model and some switching control strategies have been proposed to address a class of load balancing problems with system control input algebraic *** on the presented framework of the GSS systems and the related switching control strategies,this paper mainly studies the problem of the system control design for the GSS *** to that the GSS system is a discretely controlled continuous time system,the procedure of the control design is divided into two steps:one is to design the continuous time controller,and the other is to design and select the discrete-event driven switching control strategy.A numerical example is given to demonstrate the procedure of the system control design for the GSS systems.

关键词： System Control Design Generalized Switched Server Systems(GSS) Switching Scheduling Policy Load Balancing Hybrid Dynamic Systems(HDS) Algebraic Constraint

蝴蝶兰不同花色品种遗传多样性的RAPD分析

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上海农业学报 2003年第2期19卷 44-47页

作者：明凤董玉光娄玉霞王敬文梁斌叶鸣明沈大棱复旦大学生命科学院遗传学研究所遗传工程国家重点实验室基因多样性与设计农业中心生物多样性与生态工程教育部重点实验室上海200433 上海市农业科学院动植物引种研究中心上海201106 上海师范大学生物工程实验室上海201418

利用筛选到的 2 5个特异引物对蝴蝶兰不同花色品种作RAPD分析 ,确定了 12个品种间的遗传距离 ;并根据Neighbor joining遗传距离矩阵构建系统聚类文件 ,将该 12个品种分为两个家族 :第一家族中包含基色为白色的花色品种 ,花蕊颜色略有不... 详细信息

利用筛选到的 2 5个特异引物对蝴蝶兰不同花色品种作RAPD分析 ,确定了 12个品种间的遗传距离 ;并根据Neighbor joining遗传距离矩阵构建系统聚类文件 ,将该 12个品种分为两个家族 :第一家族中包含基色为白色的花色品种 ,花蕊颜色略有不同 ;第二家族为具有条纹的花色品种。品种间遗传距离最大为 0 .783 2 ,最小为 0 .10 3 4。

关键词：蝴蝶兰花色品种遗传多样性 RAPD分析花蕊颜色遗传距离