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Baculovirus ETL promoter acts as a shuttle promoter between insect cells and mammalian cells

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Acta Pharmacologica Sinica 2006年第3期27卷 321-327页

作者： Yu-kou LIU~(2,4) Chih-chieh CHU~(2,4) Tzong-yuan WU~(3,5) ~2Department of Chemical and Materials Engineering,Chang Gung University.Taoyuan.China:~3Department of Bioscience Technology.Chung Yuan Christian University,Chungli.China Department of Chemical and Materials Engineering Chang Gung University Taoyuan China Department of Bioscience Technology Chung Yuan Christian University Chungli China

Aim:To identify a shuttle promoter that can mediate gene expression in bothinsect cells and mammalian cells to facilitate the development of a baculovirusvector-based mammalian cell gene delivery ***:Recombinantbaculoviruses carrying the β-galactosidase reporter gene under the control of anearly to late(ETL)promoter of the Autographa californica multiple nuclear poly-hedrosis virus(AcMNPV)or a cytomegalovirus immediate early promoter(CMVpromoter)were ***1,HeLa,CHO-K1,hFobl.19,and MCF-7 mam-malian cells were tested for the expression of β-***:ETLpromoter activity was higher in bone-derived hFob 1.19 than in COS1,HeLa,CHO-K1,or MCF-7 mammalian *** transient plasmid transfection assay indi-cated that ETL promoter activity in mammalian cells was dependent on baculovirusgene ***:ETL promoter activity in mammalian cells isbaculovirus gene expression-dependent,and the shuttle promoter will facilitatethe application of baculovirus expression vectors in mammalian cell expressionsystems and for gene therapy.

关键词： baculovirus gene therapy shuttle promoter mammalian cell ETL promoter

哺乳动物细胞去核产物(胞质体及核体)的结构与功能研究

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Current Zoology 1982年第4期28卷 313-318+418-419页

作者：何大澄应建华戎宪辉丁明孝翟中和北京大学生物系

1967年Carter发现细胞松弛素可以诱发组织培养细胞的自发排核之后,Prescott(1972)借助细胞松弛素存在下的离心处理,使这一排核现象普遍化,从而确立了体外细胞去核的标准方法。经过不断改进(croce et al., 1974;Veomett et al., 1976;Luc... 详细信息

1967年Carter发现细胞松弛素可以诱发组织培养细胞的自发排核之后,Prescott(1972)借助细胞松弛素存在下的离心处理,使这一排核现象普遍化,从而确立了体外细胞去核的标准方法。经过不断改进(croce et al., 1974;Veomett et al., 1976;Lucas etal., 1976;Wigler et al., 1975),现在,这一技术已广泛应用于细胞学研究的各个重要领域(Mc Burney et al., 1979;Goldman et al., 1974;du Bols et al., 1980)。细胞去核技术及其应用的研究在我国已有初步开展(陈瑞铭等,1979;沈鼎武等,1980)。本实验对二种上皮型传代细胞系进行了去核手术,用扫描电镜和透射电镜对所获得的胞质体

关键词： mammalian cell enueleation karyopkst cytoplast function.

Research Progress on the Large-scale Culture Technology of mammalian cells

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Journal of Northeast Agricultural University(English Edition) 2009年第2期16卷 72-76页

作者： LI Chunyan XIAO Jing JIANG Yonghou The Post-doctoral Fluxion Station of Northeast Forestry University The Post-doctoral Working Station of Heilongjiang Academy of Agricultural Science Harbin 150001 China Resources and Environmental Sciences College Northeast Agricultural University Harbin 150030 China Washington National Primate Research Center University of Washington Seattle Washington 98195 USA

The culture of mammalian cells is closely related to the development of biotechnology, which has been used extensively in the research and application fields of biology and medical science. In this article, various factors affecting cell cultivation and the application of microcarrier and bioreactor on large-scale culture of mammalian ceils were reviewed.

关键词： mammalian cell culture microcarrier bioreactor

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Efficient expression of histidine-tagged large hepatitis delta antigen in baculovirus-transduced baby hamster kidney cells

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World Journal of Gastroenterology 2006年第10期12卷 1551-1557页

作者： Ying-Wei Chiang Jaw-Chin Wu Kuei-Chun Wang Chia-Wei Lai Yao-Chi Chung Yu-Chen Hu Department of Chemical Engineering National Tsing Hua University Hsinchu 300 Taiwan China Institute of Clinical Medicine National Yang Ming University and Division of Gastroenterology Department of Medicine Taipei Veterans General Hospital Taipei 112 Taiwan China

AIM： To study the baculovirus/mammalian cell system for efficient expression of functional large hepatitis delta antigen （L-HDAg）. METHODS： A recombinant baculovirus expressing histidine-tagged L-HDAg （L-HDAgH） was constructed to transduce baby hamster kidney （BHK） cells by a simplified transduction protocol. RESULTS： The recombinant baculovirus transduced BHK cells with efficiencies higher than 90% as determined by flow cytometry. The expression level was significantly higher than that obtained by plasmid transfection and was further enhanced 3-fold to around 19 pg/cell by the addition of 10 mmol/L sodium butyrate. Importantly, the expressed L-HDAgH was localized to the cell nucleus and correctly isoprenylated as determined by immunofluorescence labeling and confocal microscopy. Moreover, L-HDAgH interacted with hepatitis B surface antigen to form virus-like particles. CONCLUSION： The fusion with histidine tags as well as overexpression of L-HDAgH in the baculovirus-transduced BHK cells does not impair the biological functions. Taken together, the baculovirus/mammalian cell system offers an attractive alternative for high level expression of L-HDAgH or other proteins that require extensive posttranslational modifications.

关键词： Baculovirus Hepatitis delta virus L-HDAg mammalian cell Protein expression Transduction

TRANSCRIPTION ACTIVATION BY HBV PRE SI PROTEIN

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Journal of Pharmaceutical Analysis 1995年第2期9卷 198-198页

作者：肖生祥

L One or more hepatitis B virus(HBV)gene products may contribute to *** Pre S1 protein of HBV involves this process is not *** proteins that contain the DNA binding domain of yeast GAL4 and full-length or portions of pre S1 sequence have been used to show that the pre S1 protein contains a transcription-activating sequence that functions in yeast. The domain of pre S1 protein responsible for transcription activation is located between residues 12 and *** on transcription activation by pre S1 protein in mammalian cells are in *** preliminary results indicate that transcription activation by pre S1 protein is a possible mechanism for hepatocarcinogenesis.

关键词： HBV transcription carcinogenesis yeast mammalian cell

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