Abstract
msDNA-Ec67, a peculiar multicopy single-stranded DNA of a specific sequence was produced in NIH3T3 mouse cells. Retron-Ec67, a retroelement from Escherichia coli, was introduced under the T7 polymerase promoter and the non-translated 5'-region of the encephalomyocarditis virus. The construct was then transfected into the NIH3T3 cells constitutively producing T7 RNA polymerase. Forty-eight hours after transfection, msDNA-Ec67 was detected in the cells by means of Southern blot hybridization and reverse transcriptase extension assay. The potential use of bacterial retrons as a vector for single-stranded DNA production in mammalian cells is discussed.
Highlights
DNAof a specific sequence was produced in NIH3T3 demonstrated that msDNA-Ec67 is indeepdroduced in mouse cells
Forty-eight hours after transfection, a source of the RT coding region.An NcoI site was first introduced into msDNA-Ec67 was detected in the cells by meansof the 5'-end of the RT gene carried on YEp521-M1 by polymerase chain
A serious drawbackof t h i s technology is that the oligonucleotides have to be added from the outside of t h e cells since at present there is no method available to directly produce single-stranded DNA or oligonucleotides inside mammalian cells
Summary
(Received for publication, September 15, 1993, and in revised form, November 11, 1993). From the Department of Biochemistry, Robert Wood Johnson Medical School, Piscataway, New Jersey 08854 duced in Saccharomyces cereuisiae, and msDNA fromthis retron (msDNA-Ec67) was successfully produciendyeast cells[6]. For this purpose, retron-Ec67 was first put under controol f the T7 RNA polymerase promoter and the non-translated 5"region of the encephalomyocarditis virus (EMCV). MsDNA-Ec67, apeculiarmulticopysingle-stranded cells, constitutively expressing T7 RNA polymerase. We DNAof a specific sequence was produced in NIH3T3 demonstrated that msDNA-Ec67 is indeepdroduced in mouse cells. Erichia coli, was introduced under the T7 polymerase promoterandthenon-translated5'-region of the en-
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
