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A 3.6-kbp segment from the vir region of Ti plasmids contains genes responsible for border sequence-directed production of T region circles in E. coli

1986; Springer Nature; Volume: 5; Issue: 6 Linguagem: Inglês

10.1002/j.1460-2075.1986.tb04337.x

1460-2075

Juliane Alt-Moerbe, Bodo Rak, Joachim Schröder,

CRISPR and Genetic Engineering

Article1 June 1986free access A 3.6-kbp segment from the vir region of Ti plasmids contains genes responsible for border sequence-directed production of T region circles in E. coli Juliane Alt-Moerbe Juliane Alt-Moerbe Institut für Biologie II, Universität Freiburg, Schänzlestrasse 1, D-7800 Freiburg, FRG Search for more papers by this author Bodo Rak Bodo Rak Institut für Biologie III, Universität Freiburg, Schänzlestrasse 1, D-7800 Freiburg, FRG Search for more papers by this author Joachim Schröder Joachim Schröder Institut für Biologie II, Universität Freiburg, Schänzlestrasse 1, D-7800 Freiburg, FRG Search for more papers by this author Juliane Alt-Moerbe Juliane Alt-Moerbe Institut für Biologie II, Universität Freiburg, Schänzlestrasse 1, D-7800 Freiburg, FRG Search for more papers by this author Bodo Rak Bodo Rak Institut für Biologie III, Universität Freiburg, Schänzlestrasse 1, D-7800 Freiburg, FRG Search for more papers by this author Joachim Schröder Joachim Schröder Institut für Biologie II, Universität Freiburg, Schänzlestrasse 1, D-7800 Freiburg, FRG Search for more papers by this author Author Information Juliane Alt-Moerbe1, Bodo Rak2 and Joachim Schröder1 1Institut für Biologie II, Universität Freiburg, Schänzlestrasse 1, D-7800 Freiburg, FRG 2Institut für Biologie III, Universität Freiburg, Schänzlestrasse 1, D-7800 Freiburg, FRG The EMBO Journal (1986)5:1129-1135https://doi.org/10.1002/j.1460-2075.1986.tb04337.x PDFDownload PDF of article text and main figures. ToolsAdd to favoritesDownload CitationsTrack CitationsPermissions ShareFacebookTwitterLinked InMendeleyWechatReddit Figures & Info The vir region of Ti plasmids is responsible for the transfer of the T region from Agrobacteria to plant cells; previous experiments suggested that formation of independent T region DNA circles is one step in this process. To study this step in Escherichia coli, we developed a binary vector system. One plasmid (=substrate) contains correctly oriented right and left borders from octopine plasmid pTiAch5. A gene with a counterselectable function (galK) was cloned between these borders. The galK gene is under the control of the tac promoter–operator and the lac repressor with the laci gene also in the selection cassette. This construction allows determination of substrate plasmid mutants which have lost the selectable galK function. The second component of the system is one of a set of compatible plasmids harbouring various cloned parts from the vir region of nopaline plasmid pTiC58. A 3.6-kbp segment of the vir region turned out to be necessary and sufficient for production of substrate plasmid mutants which represented the equivalent of the T region containing a complete left border. From this vir region fragment four discrete proteins were expressed in minicells. The coding regions were mapped to a part conserved in nopaline and octopine plasmids; in the latter it appears to correspond to virC/D. Previous ArticleNext Article Volume 5Issue 61 June 1986In this issue RelatedDetailsLoading ...