Plasmid rescue from transgenic mouse DNA using LacI repressor protein conjugated to magnetic beads

Autor:	J A, Gossen, W J, de Leeuw, A C, Molijn, J, Vijg
Rok vydání:	1993
Předmět:	Escherichia coli Proteins Genetic Vectors DNA Recombinant Mice Transgenic Bacteriophage lambda Repressor Proteins Magnetics Mice Bacterial Proteins Genetic Techniques Lac Operon Evaluation Studies as Topic Escherichia coli Lac Repressors Animals Biotechnology Plasmids
Zdroj:	BioTechniques. 14(4)
ISSN:	0736-6205
Popis:	A method for the efficient rescue of lac operator containing plasmids from transgenic mouse genomic DNA is described. The method is based on the high affinity of the LacI repressor protein for the lac operator sequence. Using the LacI repressor protein conjugated to magnetic beads, more than 95% of plasmid sequences could be purified from restriction enzyme digested genomic DNA. After circularization, the plasmids were introduced into Escherichia coli by means of electroporation. Since the plasmid was cloned into a bacteriophage lambda vector, the efficiency of plasmid rescue could easily be compared with in vitro packaging. Our results indicate that plasmid rescue is about 25 times more efficient. Application of this method should be especially useful with transgenic mouse models harboring LacZ plasmid shuttle vectors for studying spontaneous or induced mutations in vivo.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=pmid________::2dadb9cd6353433ad10bbbeab8bc0a6d https://pubmed.ncbi.nlm.nih.gov/8476606 Zobrazit plný text záznamu