EXPRESSION OF HEPATITIS B VIRUS GENES IN TISSUE CULTURE CELLS.
Item
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Title
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EXPRESSION OF HEPATITIS B VIRUS GENES IN TISSUE CULTURE CELLS.
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Identifier
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AAI8708329
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identifier
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8708329
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Creator
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ZELENT, ARTHUR ZYGINANT.
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Contributor
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George Acs
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Date
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1987
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Language
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English
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Publisher
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City University of New York.
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Subject
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Biology, Molecular
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Abstract
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In order to study the life cycle of hepatitis B virus (HBV) our laboratory attempted to develop a tissue culture system which would support the viral replication. In the course of this study two cell lines were developed which markedly differed with respect to HBV gene expression.;The 4.10 cell line was established by cotransfection of mouse 3T3 cells with plasmid DNA containing a head-to-tail dimer of the HBV genome and DNA coding for methotrexate resistant dihydrofolate reductase. These cells contain at least 40 copies of intact HBV dimer per cell, and produce large amounts of 22 nm hepatitis B surface antigen (HBsAg) particles that include viral major and middle envelope proteins. They also synthesize viral "e" antigen (HBeAg). However, the presence of core antigen (HBcAg) was not detected.;Analysis of mRNAs isolated from 4.10 cells revealed the presence of two major HBV transcripts the 2.1 kilobase (kb) mRNA and 3.5 kb longer-than-genome length RNA , although the amount of the latter species was approximately 0.1% of the former. In contrast to this cell line, high levels of the longer-than-genome length RNA and HBcAg expression were observed in D1113 cells. These cells were established by transfecting Moloney murine leukemia virus retroviral vector (carrying two head-to-tail HBV dimers and gene coding for neomycin resistance) into a transformed mouse fibroblast cell line ((psi)AM22b). The data presented here represents the first example of a tissue culture system in which the two major HBV mRNAs are expressed in relatively equal amounts similar to the proportions observed during in vivo infection. The initiation sites for the 2.1 kb and the 3.5 mRNAs were mapped 5' to and within the pre-S2 region and 5' to and within the pre-C region of HBV, respectively. These positions correspond very well with those from which the same mRNAs are transcribed in infected livers. Since the D1113 cells produce abundant amounts of the most important marker of HBV replication, the 3.5 kb "pre-genome" RNA, they should have a potential to support at least partial replication of the virus and assembly of "Dane-like" particles.
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Type
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dissertation
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Source
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PQT Legacy CUNY.xlsx
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degree
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Ph.D.
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Program
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Biomedical Sciences
