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Volume 63, Issue 1

Molecular Cloning of the Endogenous Rat C-type Helper Virus DNA Sequence: Structural Organization and Functional Analysis of Some Restricted DNA Fragments Free

Abstract

SUMMARY

Recently, we have identified and purified the integrated and proviral sequences specific for two endogenous rat type C leukaemia helper viruses: which originated from a fibrosarcoma induced in a feral rat and from the cell line which originated from a Buffalo rat hepatoma. The rat leukaemia helper virus sequences have previously been shown to be 8.4 to 8.8 kilobases (kb) in size. In this communication, we report the molecular cloning of the 8.8 kb of by ligation at the site of the vector p322, cultured in an host. After screening 5750 clones for ampicillin resistance and tetracycline sensitivity and testing by colony hybridization using P-labelled c, four clones were isolated, two of which carried the total 8.8 kb . A detailed restriction endonuclease map of the cloned was deduced by sequential digestions of either 3′- or 5′-labelled . Of the 14 restriction enzymes tested, RI, HI, I, I, I, II and I gave informative cleavage patterns. At least two copies of long terminal repeated sequences () flanking the 3′ and 5′ termini of the proviral were identified by I and I cleavages. in the rat endogenous leukaemia helper virus measured 780 ± 20 nucleotides in length. The genetic information encoded by the cloned was also analysed by hybridization selection of m, which was then used in cell-free protein synthesis in a rabbit reticulocyte lysate system. Essentially all major RaLV-specific proteins precipitable by anti-RaLV serum were synthesized , confirming that the genomic was successfully cloned with little fidelity loss or scrambling of the genetic information.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): DNA cloning , hybrid-select translation , rat retrovirus and restriction map
© Society for General Microbiology 1982
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1982-11-01
2024-04-25
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Molecular Cloning of the Endogenous Rat C-type Helper Virus DNA Sequence: Structural Organization and Functional Analysis of Some Restricted DNA Fragments
J. Gen. Virol. 63, 25 (1982); https://doi.org/10.1099/0022-1317-63-1-25
/content/journal/jgv/10.1099/0022-1317-63-1-25
/content/journal/jgv/10.1099/0022-1317-63-1-25
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