Restriction enzyme-mediated integration elevates transformation frequency and enables co-transfection of Toxoplasma gondii (Record no. 40457)

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Transcribing agency CICY
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Classification number (OCLC) (R) ; Classification number, CALL (RLIN) (NR) B-6112
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Title Restriction enzyme-mediated integration elevates transformation frequency and enables co-transfection of Toxoplasma gondii
490 0# - SERIES STATEMENT
Volume/sequential designation Molecular and BioChemical Parasitology, 74(1), p.55-63, 1995
520 3# - SUMMARY, ETC.
Summary, etc. This report describes the use of restriction enzyme-mediated integration (REMI)to increase the transformation frequency and allow co-transfection of several unselected constructs under the selection of a single selectable marker. We found that while BamHI (the enzyme used to originally demonstrate REMI (Schiestl, R.H. and Petes, T.D. (1991)Integration of DNA fragments by illegitimate recombination in Saccharomyces cerevisiae. Proc. Natl. Acad. Sci. USA 88, 7585-7589)increased the number of transformants by 2-5-fold over the control without added enzyme, NotI proved to be a further 29-46-times more effective in enhancing stable transformation. This simple technique was used in the transformation of three non-selective markers (two modified membrane proteins and -galactosidase)with a selectable construct expressing chloramphenicol acetyltransferase. Following chloramphenicol selection, four out of ten independent transformants stably acquired all four constructs with at least two expressing all four genes at the protein level. These results demonstrate that REMI may be used in the efficient stable transformation and co-transfection of this and perhaps other protozoan parasites.
650 14 - SUBJECT ADDED ENTRY--TOPICAL TERM
Topical term or geographic name entry element RESTRICTION ENZYME-MEDIATED INTEGRATION (REMI)
650 14 - SUBJECT ADDED ENTRY--TOPICAL TERM
Topical term or geographic name entry element NOTI
650 14 - SUBJECT ADDED ENTRY--TOPICAL TERM
Topical term or geographic name entry element BAMHI
650 14 - SUBJECT ADDED ENTRY--TOPICAL TERM
Topical term or geographic name entry element CO-TRANSFECTION
650 14 - SUBJECT ADDED ENTRY--TOPICAL TERM
Topical term or geographic name entry element TRANSFORMATION
700 12 - ADDED ENTRY--PERSONAL NAME
Personal name Black, M.
700 12 - ADDED ENTRY--PERSONAL NAME
Personal name Seeber, F.
700 12 - ADDED ENTRY--PERSONAL NAME
Personal name Soldati, D.
700 12 - ADDED ENTRY--PERSONAL NAME
Personal name Kim, K.
700 12 - ADDED ENTRY--PERSONAL NAME
Personal name Boothroyd, J.C.
856 40 - ELECTRONIC LOCATION AND ACCESS
Uniform Resource Identifier <a href="https://drive.google.com/file/d/1hT3HYo-KJMDPmz7wO7myNYjTCx0kxBJf/view?usp=drivesdk">https://drive.google.com/file/d/1hT3HYo-KJMDPmz7wO7myNYjTCx0kxBJf/view?usp=drivesdk</a>
Public note Para ver el documento ingresa a Google con tu cuenta: @cicy.edu.mx
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  Clasificación local     Ref1 CICY CICY Documento préstamo interbibliotecario 25.06.2025   B-6112 25.06.2025 25.06.2025 Documentos solicitados