Isolation of recombinant DNA clones carrying complete integrated proviruses of Moloney murine leukemia virus

Bacheler, Lee T.; Fan, Hung

doi:10.1128/jvi.37.1.181-190.1981

Cited by 52 publications

(22 citation statements)

References 30 publications

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“…In addition, pre-S(1) sequences were detected on the secreted HBsAg. As each large HBsAg polypeptide contains the pre-S(2) sequence in addition to the pre-S (1) sequence, it is likely that this secreted HBsAg contains no more than 5% pre-S(1)-containing molecules.…”

Section: Resultsmentioning

confidence: 99%

Expression of hepatitis B virus surface and core antigens: influences of pre-S and precore sequences

McLachlan¹,

Milich²,

Raney³

et al. 1987

J Virol

163

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Amphotropic retroviral expression systems were used to synthesize hepatitis B virus surface antigen (HBsAg) and core antigen. The vectors permitted establishment of cell lines which expressed antigen from either the retroviral long terminal repeat or the mouse metallothionein-I promoter. HBsAgs were synthesized containing no pre-S sequences, pre-S(2) sequences alone, or pre-S(l) plus pre-S(2) sequences. Inclusion of pre-S(2) sequences did not affect the secretion or density of HBsAg particles but did reduce their mass by approximately 30%. Addition of pre-S(l) sequences almost completely abolished secretion of HBsAg and resulted in its localization in an aqueous-nonextractable preor early-Golgi cellular compartment. HBsAg was localized to the cytoplasm of the cell. This localization was unaffected by the presence of pre-S sequences in the antigen. Cell lines synthesizing hepatitis B antigens from core DNA fragments, containing or not containing precore sequences, secreted hepatitis B e antigen. However, the absence of precore DNA sequences resulted in additional synthesis of hepatitis core antigen, which was predominantly nuclear in localization.

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Section: Resultsmentioning

confidence: 99%

Expression of hepatitis B virus surface and core antigens: influences of pre-S and precore sequences

McLachlan¹,

Milich²,

Raney³

et al. 1987

J Virol

163

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“…The proviral Mo-MuLV DNA used in this work originates from clone #48, which is a lambda clone containing an EcoRI DNA fragment from a Mo-MuLV infected mouse fibroblast line (14). This DNA fragment was subcloned into EcoRI cut pBR 322.…”

Section: Materials and Methods Materialsmentioning

confidence: 99%

The effect of specific mutations at and around thegag-polgene junction of Moloney murine leukaemia virus

Jones¹,

Nemoto²,

Kuchino³

et al. 1989

Nucl Acids Res

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By carrying out oligonucleotide-directed mutagenesis, in vitro, on a 3.3 kb XhoI-HindIII fragment from Moloney murine leukaemia virus Mo-MuLV proviral DNA, inserted into the phagemid pTZ19R, nine separate fragments have been prepared in which mutations have been inserted at and around the gag-pol gene junction. Using these mutant fragments Mo-MuLV proviral DNA has been reassembled and cloned into pBR322. Examination of the mutant proviral DNAs in mouse culture cells indicates that a terminator codon at the gag-pol junction is essential for function, but any of the three chain terminator codons gives an active virus. Also the region of secondary structure surrounding the terminator codon must be preserved.

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“…This cDNA had a specific activity of 1 x 108 to S x 108 cpm/,ug. A selected, AKR-MuLV-specific cDNA probe was obtained by pre-hybridization to a 1,000-fold excess of M-MuLV virion RNA (1,46).…”

Section: Methodsmentioning

confidence: 99%

DNA methylation affecting the expression of murine leukemia proviruses

Hoffmann¹,

Steffen²,

Gusella³

et al. 1982

J Virol

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The endogenous, vertically transmitted proviral DNAs of the ecotropic murine leukemia virus in AKR embryo fibroblasts were found to be hypermethylated relative to exogenous AKR murine leukemia virus proviral DNAs acquired by infection of the same cells. The hypermethylated state of the endogenous AKR murine leukemia virus proviruses in these cells correlated with the failure to express AKR murine leukemia virus and the lack of infectivity of cellular DNA. Induction of the endogenous AKR murine leukemia virus proviruses with the methylation antagonist 5-azacytidine suggested a causal connection between DNA methylation and provirus expression. Also found to be relatively hypermethylated and noninfectious were three of six Moloney murine leukemia virus proviral DNAs in an unusual clone of infected rat cells. Recombinant DNA clones which derived from a methylated, noninfectious Moloney provirus of this cell line were found to be highly active upon transfection, suggesting that a potentially active proviral genome can be rendered inactive by cellular DNA methylation. In contrast, in vitro methylation with the bacterial methylases MHpaII and MHhaI only slightly reduced the infectivity of the biologically active cloned proviral DNA. Recombinant DNA clones which derived from a second Moloney provirus of this cell line were noninfectious. An in vitro recombination method was utilized in mapping studies to show that this lack of infectivity was governed by mechanisms other than methylation.

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Isolation of recombinant DNA clones carrying complete integrated proviruses of Moloney murine leukemia virus

Cited by 52 publications

References 30 publications

Expression of hepatitis B virus surface and core antigens: influences of pre-S and precore sequences

Expression of hepatitis B virus surface and core antigens: influences of pre-S and precore sequences

The effect of specific mutations at and around thegag-polgene junction of Moloney murine leukaemia virus

DNA methylation affecting the expression of murine leukemia proviruses

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