Modified blasticidin S resistance gene (bsrm) as a selectable marker for construction of retroviral vectors

Modified blasticidin S resistance gene (bsrm) as a selectable marker for construction of retroviral vectors

Autor Freitas, A. C. Google Scholar
Bento, F. M. Google Scholar
Ramesh, N. Google Scholar
Osborne, WRA Google Scholar
Han, S. W. Google Scholar
Instituição Universidade Federal de São Paulo (UNIFESP)
Univ Washington
Resumo Retroviral vectors are commonly used in ex vivo gene therapy protocols. the structure of vectors basically consists of one gene of interest and a selectable marker gene. Fast selection without damaging cells is a critical step for ex vivo gene therapy protocols. Blasticidin S deaminase isolated from Bacillus cereus has a neutralizing action on the highly toxic antibiotic blasticidin S (BS). A commercially available gene coding for blasticidin S deaminase (bsr) when used to construct retroviral vectors, LBSN and LNSB, provided very low levels of BS deaminase activity, precluding their routine use in gene transfer experiments. However, with the introduction of specific mutations into the bsr gene based on the Kozak consensus sequences and deletion of a 5' untranslated sequence to generate bsrm, we were able to construct a retroviral vector encoding resistance to high doses of BS (at least 16-fold above the usual lethal dose in NIH3T3 cells), showing that bsrm/BS may provide a useful system for selection of transduced mammalian cells. (C) 2002 Elsevier Science B.V. All rights reserved.
Palavra-chave bsr
selectable marker
retroviral vector
mutagenesis
blasticidin S deaminase
Idioma Inglês
Data de publicação 2002-04-25
Publicado em Journal of Biotechnology. Amsterdam: Elsevier B.V., v. 95, n. 1, p. 57-62, 2002.
ISSN 0168-1656 (Sherpa/Romeo, fator de impacto)
Publicador Elsevier B.V.
Extensão 57-62
Fonte http://dx.doi.org/10.1016/S0168-1656(01)00442-4
Direito de acesso Acesso restrito
Tipo Artigo
Web of Science WOS:000174859800006
Endereço permanente http://repositorio.unifesp.br/handle/11600/26834

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