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Chinese Science Bulletin 1999年第2期44卷 158-163页

作者： Ying Zhu yipeng qi Wuhan Univ Inst Virol Wuhan 430072 Peoples R China

A transposon-shuttle vector Hanpvid was constructed by using wild-type genomic DNA from Heliothis armigera nuclear polyhedrosis virus (HaNPV). lt could replicate in E. coli cells as a large plasmid and remain infectious when being induced into insect *** comprises HaNPV DNA and a transposon cassette which includes a miniF replicon, a kanamycin resistance gene (kan), lacZa and an attachment site for Tn7 (attTn7). Recombinant virus rHa-FaGP was obtained after transposition of a donor plasmid carrying green fluorescent protein gene (gfp) and polyhedrin gene (ocu) into ***-PAGE analysis shows that both gfp and ocu genes were highly expressed in Heliothis armigera cells. Green Hemolymphocytes can be seen under a fluorescent microscope 4 d after recombinant virus rHa-FaGP infected the third-instar larvae. The infected larvae show strong green fluorescence 6 d post infection.

关键词： HaNPV transposon-shuttle vector recombinant virus green fluorescent protein.

Influence of coronary bifurcation angle on atherosclerosis

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Acta Mechanica Sinica 2019年第6期35卷 1269-1278页

作者： Zhaomiao Liu Shengwei Zhao Yunjie Li Feng Shen yipeng qi qi Wang College of Mechanical Engineering and Applied Electronics Technology Beijing University of TechnologyBeijing 100124China Department of Cardiology PLA General HospitalBeijing 100853China

Hemodynamics plays a crucial role in the development and progression of coronary atherosclerosis,which is prone to occur in branch bifurcation.A n individual aortic-coronary artery model and three changed bifurcation angle models are constructed by Mimics and Freeform based on computed tomography *** influence of different coronary bifurcation angles between left main(LM),left anterior descending(LAD),and left circumflex(LCX)on the blood flow field and related hemodynamic parameters are *** is shown that a wider bifurcation angle between LAD and LCX can cause a wider low-wall shear stress area,leading to ***,a decreased angle between LM and LAD is predisposed to prevent *** results help to better understand the hemodynamic causes of atherosclerosis with various bifurcation angles in coronary arteries and to provide guidance for clinical assessment and prevention.

关键词： Atherosclerosis Coronary artery Bifurcation angle Hemodynamics Wall shear stress

Measles Virus(Nepal Strain)Hemagglutinin Gene: Cloning,Complete Nucleotide Sequence Analysis and Expression in COS Cells

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Wuhan University Journal of Natural Sciences 1998年第3期3卷 123-128页

作者： Li, Lingyun qi, yipeng College of Life Sciences Wuhan University Wuhan China

Hemagglutinin gene of Measles virus(Nepal strain) was amplified by RT PCR technique, cloned and sequenced by the dideoxy mediated chain termination method. The comparison to the standard strain(Edmonston strain) showed many important mutations. The homology of these two strains was 98.17%. Then H gene was cloned into expression vector pCD SRα296 and introduced into COS 7 cells by electroporation method. The expression and function of cloned H gene was checked by hemadsorption assays.

关键词： measles virus hemagglutinin gene,cos 7 cells,hemadsorption assays

Structure and function of bak gene and its apoptosis-inducing mechanism

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Chinese Science Bulletin 1999年第7期44卷 605-609页

作者： Yan Li yipeng qi Gengfu Xiao Wuhan Univ Inst Virol Wuhan 430072 Peoples R China

The Bcl-2 family plays an important role in the regulation of apoptosis. Some members in the family prevent apoptosis while others promote it. The newly discovered bcl-2 homolog,bak, promotes the apoptosis induced by a variety of stimuli. It can interact with the anti-apoptotic members through its BH3 domain and antagonize their activity. Investigation results suggest that the expression of bak gene becomes altered in some diseases, such as cancer. The preferential stimulation of Bak-induced apoptosis, therefore, may be involved in the mechanisms of some antitumor drugs.

关键词： programmed cell death apoptosis Bcl-2 family bak.

Construction and function of recombinant AcMNPV with double copies of v-cath gene

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Chinese Science Bulletin 2002年第1期47卷 52-57页

作者： Deli Liu Huazhong Xiao yipeng qi Lunguang Yao Wuhan Univ Inst Virol Wuhan 430072 Peoples R China

Two recombinant baculoviruses, dciAcMNPV and dcdAcMNPV in which another copy of the v-cath gene controlled by ie1 promoter and polh promoter was inserted, were respectively constructed by the Bac-to-Bac system. The expression of the v-cath gene of the recombinant baculovi-ruses in Sf9 cells at different phases was investigated by SDS-PAGE and Western blot. The results showed that only re-combinant virus dciAcMNPV containing late gene v-cath driven by early gene promoter could express V-CATH pro-tein, cathepsin encoded by virus genome, 12 h post-infection and dcdAcMNPV containing late gene v-cath driven by late and very late gene promoters could express more V-CATH protein. Negative control ncAcMNPV, a mutant deleted v-cath gene, could not express V-CATH protein at all. The Spodopera exigua larvae were infected with viruses respec-tively and the results showed that the toxicity was as follows: dcdAcMNPV>dciAcMNPV>wtAcMNPV>ncAcMNPV. The toxicity of recombinant viruses and the characters of dead larvae

关键词： v-cath gene Western blot Bac-to-Bac system recombi-nant baculovirus.

asy and asyip:A new type of apoptosis-inducing gene

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Chinese Science Bulletin 2001年第17期46卷 1409-1411页

作者： Bing qi yipeng qi qinzheng Liu Xiaoling Qu Wuhan Univ Inst Virol Wuhan 430072 Peoples R China

Apoptosis,,a wide physiological process in multicel-lular organisms, is critical for the organ development, the cell senescence, the tissue homeostasis and the resistance to infection of virus and bacteria. Apoptosis plays a key role in the regulation of the growth cell number such as the tissue construction, the elimination of abnormal or dangerous cells. Therefore, apoptosis is a stringent and effective cell quality controlling system, which reduces the number of harmful cells to the maximum, such as auto-immunity cell, cells infected by virus, tumor cells by cell suicide111. Apoptosis can be initiated by a wide variety of the extracellular and intracellular stimuli, including the developmental signals, the cellular stress and the disruption of cell cycle, transduced and amplified by the second messengers, and finally finished by the activating death effector protease. This process can be called apoptosis signal network. The defective in control of the apoptotic pathways may contribute to a variety

关键词： gene cell type

Analysis of proteins that interact with nucleocapsid protein of SARS-CoV using 15-mer phage-displayed library

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Chinese Science Bulletin 2007年第15期52卷 2072-2080页

作者： LIU ZhengXue WANG ZhanHui LIU YingLe DONG Wei qi yipeng State Key Laboratory of Virology College of Life Science Wuhan University Wuhan 430072 China Biology Department Chongqing Three Gorges University Chongqing 404000 China

Analysis of proteins that interact with N protein of SARS-CoV using 15-mer phage-displayed library will help to explore the virus pathogenesis and to develop new drugs and vaccines against SARS. In this study,we cloned,expressed and purified N protein of SARS-CoV. This 46-kD N protein was verified by SDS-PAGE and Western-blot. Then,the peptides binding-specific to N protein were identified using 15-mer phage-displayed library. Surprisingly,all of the 89 clones from monoclonal ELISA were positive (S/N>2.1) and the result was further confirmed experimentally once again. Six N protein-binding pep-tides,designated separately as SNA1,SNA2,SNA4,SNA5,SNA9 and SNG11,were selected for se-quencing. Sequence analysis suggested that SNA5 shared approximatively 100% sequence identity to SNA4,SNA2,SNA9 and SNA1. In addition,the binding specificity of the 15-mer peptides with the SARS-CoV N protein was further demonstrated by blocking ELISA using the synthetical 15-mer peptide according to the deduced amino acid sequence of SNA5. Also,the deduced amino sequence of SNA5 was compared with proteins in translated database using the tblastx program,and the results showed that the proteins with the highest homology were Ubiquinol-cytochrome c reductase iron-sulfur sub-units (UCRI or UQCR),otherwise known as the Rieske iron-sulfur proteins (RISP). Notablely,in the 2Fe-2S redox centre of UCRI,there were 6 residues GGW(Y)F(Y)CP compatible to the residues (po-sition 2→7,GGWFCP7) of the NH2-terminal of the 15-mer peptide,which indicated higher binding specificity between the N protein of SARS-CoV and the redox centre of UCRI to some extent. Here,the possible molecular mechanisms of SARS-CoV N protein in the pathogenesis of SARS are discussed.

关键词：非典型肺炎核蛋白壳抗菌素细胞凋亡发病机制

Cloning and identification of measles virus receptor gene from marmoset cells

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Chinese Science Bulletin 2002年第20期47卷 1679-1687页

作者： Lingyun Li Xin Liu Peng Zhang yipeng qi Ming Cheng Wuhan Univ Inst Virol Wuhan 430072 Peoples R China

The measles virus (MV) strains with mutated hemagglutinin gene (ha) lost the capacity to infect its sensitive host cells (Vero cells), but it may infect the marmoset B-lymphoblastoid cell line B95a. From above, we can presume that there is a novel cellular receptor for those measles virus strains on B95a cells. Using the yeast two-hybrid system, we screened and cloned a novel gene-bip (B-lymphoblastoid interaction protein of marmoset) from B95a cell cDNA library, which encoded a protein interacting with measles virus hemagglutinin protein (Ha). The bip cDNA was 1540 base pairs in length and contained a unique open reading frame (ORF) of 1011 base pairs encoding a transmembrane protein of 337 amino acid residues. The primary structure of amino acids residue is predicted that the Bip comprised a hydrophobic transmembrane domain and a hydrophobic leader region. The researches about the deletion mutants showed that the deletion of transmembrane domain in Bip did not affect the interaction between Bip and

关键词： B95a-derived measles virus isolates hemagglutinin pro-tein (Ha) yeast two-hybrid system hip slam.

COLIPHAGE M13 CLONING SYSTEM AND ddXTP CHAIN-TERMINATION METHOD FOR DNA SEQUENCING

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中国科学B辑(英文版) 1986年第6期29卷 609-617页

作者：齐义鹏 Department of Virology Wuhan University

Two DNA fragments of cytochrome b gene in yeast mitochondral DNA were sequenced by Messing’s M13 cloning system and Sanger’s ddXTP chain-termination method. M13mp8 and M13mp9 serve as vectors for insert fragment. The recipient strain is E. coli JM103 for transfection. When the ratio of insert/vector was 3:1, high frequencies of recombination and positive recombination were obtained. The two fragments, which have 575 bp and 709 bp, were sequenced. To read more bases, the ratio of ddXTP/dXTP must be reduced. On one X-ray film of 80×17cm 394 bp were read.

关键词： sequenced recombination cloning DNA Sanger insert transfection yeast recipient restr

Cloning and Expression of HIV-1 p24 Gene in Insect Cells by Using BAC-TO-BAC System

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Wuhan University Journal of Natural Sciences 1998年第1期3卷 113-118页

作者： Mallam Nock Joshua qi yipeng Huang Yongxiu Liu Ziye College of Life Sciences Wuhan University Wuhan China

Recombinant transposing vector pFHIV24 was constructed by cloning the HIV-1 p24 gene into the Multiple cloning site (MCS) of the transposing vector pFastBac1 in the correct orientation with respect to the polyhedrin promoter. Recombinant bacmid bHIV24 was obtained by transposing a mini-att Tn7 element from the recombinant pFHIV24 to the mini-att Tn7 attachment site on the bacmid by Tn7 transposition functions provided by the helper plasmid. Minipreparation of recombinant bacmid DNA was transfected intoSpodoptera frugiperda (Sf9) cells to get the recombinant virus. Fresh insect Sf9 cells were infected with the recombinant virus containing p24 to express the target protein. The target protein expressed was analyzed on a 15% polyacrylamide gels and then used as antigen to check HIV-1 positive serum by ELISA. Our positive result shows that the expressed p24 protein could be used as standard antigen for HIV-1 diagnosis by ELISA and other reliable diagnostic methods of HIV-1 infection.

关键词： cloning expression baculovirus antigen antibody ELISA