Recombineering BAC transgenes for protein tagging.
Methods
; 53(2): 113-9, 2011 Feb.
Article
em En
| MEDLINE
| ID: mdl-20868752
ABSTRACT
Protein tagging offers many advantages for proteomic and regulomic research, particularly due to the use of generic and highly sensitive methods that can be applied with reasonable throughput. Ideally, protein tagging is equivalent to having a high affinity antibody for every chosen protein. However, these advantages are compromised if the tagged protein is overexpressed, which is usually the case from cDNA expression vectors. BAC (bacterial artificial chromosome) transgenes present a way to express a chosen protein at physiological levels with all regulatory elements in their native configurations, including cell cycle, alternative splicing and microRNA regulation. Recombineering has become the method of choice for modifying large constructs like BACs. Here, we present a method for protein tagging by recombineering BACs, transfecting cells and evaluating tagged protein expression.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Transgenes
/
Cromossomos Artificiais Bacterianos
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Proteômica
Limite:
Animals
Idioma:
En
Ano de publicação:
2011
Tipo de documento:
Article