Experimental manipulation of gene expression / edited by Masayori Inouye.

Date:
1983
Catalogue details

Licence: Attribution-NonCommercial 4.0 International (CC BY-NC 4.0)

Credit: Experimental manipulation of gene expression / edited by Masayori Inouye. Source: Wellcome Collection.

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    8. Construction of Cloning Vehicles from Murine Retroviruses 161 EcoRI ^Hindm Mink-cell DNA EcoRI HindM Mink-Cell DNA ì\ EcoRI Psfl 2490 BarnHI noo Deleted Segment ЕсоП1--ЕСОН /1 Polyoma ori and ^ Early region Hpol A Ecogpt BamHI Fig.3 Construction of pMuSV-g/7i. The starting materials for this construction are shown at the top of the figure; plM, a hybrid plasmid DNA containing a mink cell DNA-derived integrated copy of the Ml M-MuSV viral genome (see Van de Woude et al., 1979) and a modified pBR322 genome that lacks poison sequences, which have been shown to inhibit replication of plasmid recombinants in mammalian cells (Lusky and Botchan, 1981). Through the series of enzymatic manipulations out¬ lined, the final product, pMuSV-gpi, was constructed. In pMuSV-gpi, the direction of viral transcrip¬ tion is counterclockwise, as is the direction of transcription of the polyoma virus early region. The boxed long terminal repeat (LTR) and v-mos sequences denote the long terminal redundancies found in retrovirus proviral DNA and MSV transforming sequences, respectively. The Pst\ site in the pMuSV molecule is located ~50 nucleotides to the 5' side of the AUG codon initiating gag-pol translation (Shinnick et al., 1981). The actual details of the construction are presented elsewhere (R. C. Mulligan, unpublished).