Methods for single/low-copy integration by ultraviolet and trimethylpsoralen treatment in Caenorhabditis elegans.
Methods
; 68(3): 397-402, 2014 Aug 01.
Article
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| MEDLINE
| ID: mdl-24613935
ABSTRACT
Single/low-copy transgene integration is essential for avoiding overexpression, ectopic expression and gene silencing in the germline. Here, we present an overview of a method that uses ultraviolet and trimethylpsoralen (UV/TMP) to generate single/low-copy gene integrations in Caenorhabditis elegans. Single/low-copy transgenes from extrachromosomal arrays are integrated into the genome using positive selection based on temperature sensitivity with a vps-45 rescue fragment and negative selection based on benzimidazole sensitivity with a ben-1 rescue fragment. The copy number of the integrated transgenes is determined using quantitative PCR. Our UV/TMP integration method, which is based on familiar extrachromosomal transgenics, provides a simple approach for generating single/low-copy gene integrations.
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Texto completo:
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Bases de datos:
MEDLINE
Asunto principal:
Animales Modificados Genéticamente
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Caenorhabditis elegans
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Transgenes
Límite:
Animals
Idioma:
En
Revista:
Methods
Asunto de la revista:
BIOQUIMICA
Año:
2014
Tipo del documento:
Article