Efficient construction of plant genomic libraries requires the use ofmcr host strains and packaging mixes

Autor:	David M. Woodcock, Michael W. Graham, J.P. Doherty
Rok vydání:	1990
Předmět:	Genetics Cloning medicine.medical_specialty Host (biology) food and beverages Plant Science Genomic cloning Biology chemistry.chemical_compound 5-Methylcytosine chemistry Molecular genetics medicine Genomic library Molecular Biology Gene DNA
Zdroj:	Plant Molecular Biology Reporter. 8:18-27
ISSN:	1572-9818 0735-9640
DOI:	10.1007/bf02668877
Popis:	It has recently become apparent that many strains ofE. coli contain nucleases encoded by themcrA andmcrB loci that, recognize the modified base 5-methylcytosine in DNA. Plant DNAs have particularly high levels of this modification and the activity of these 5-methylcytosine-specific nucleases is particularly relevant to cloning plant genomic DNAs. We show here that for preparing libraries in a λ replacement vector, the use of suitablemcr hosts andmcr packaging mixes can increase the efficiency of cloning of plant genomic DNAs by at least two orders of magnitude. We also provide evidence that the activity of themcr nucleases is probably a significant source of bias in the representation of sequences in plant genomic libraries.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_________::39c9b4f7b15490189fe900a96ca35ae3 https://doi.org/10.1007/bf02668877 Zobrazit plný text záznamu Plný text ve formátu PDF