Autor:	Dulce M. Santos, Ana Sofia Duque, Manuel Pedro Salema Fevereiro, Susana de Sousa Araújo
Rok vydání:	2003
Předmět:	Genetics Agrobacterium fungi Chimeric gene Agrobacterium tumefaciens Horticulture Biology biology.organism_classification Molecular biology Reverse transcription polymerase chain reaction Transformation (genetics) Plasmid Complementary DNA Gene
Zdroj:	Plant Cell, Tissue and Organ Culture. 74:293-296
ISSN:	0167-6857
DOI:	10.1023/a:1024093105154
Popis:	We have developed a rapid and simple RT-PCR based method to check the integrity of chimeric genes within plasmid constructs for plant transformation. It exploits the Agrobacterium tumefaciens-mediated transient expression of plasmid constructs in plant tissue. Total RNA was isolated from tobacco leaves co-cultivated for 3 days with Agrobacterium tumefaciens harbouring the plant transformation vectors constructed in our laboratory. First strand cDNA synthesis with oligo(dT) primers generate a pool of cDNA that was used for PCR amplification with primers specific for each of the genes present within the constructs. PCR amplification reactions were successful for all chimeric genes tested, thus confirming their intactness and suitability to be used for stable plant transformation.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_________::b7ed31da7f1d4152f29a0708b0fabcc8 https://doi.org/10.1023/a:1024093105154 Zobrazit plný text záznamu Full text from SpringerLink