Genetic transfer and expression of reconstructed yeast artificial chromosomes containing normal and translocated BCL2 proto-oncogenes

Autor:	G A Silverman, E Yang, J H Proffitt, M Zutter, S J Korsmeyer
Rok vydání:	1993
Předmět:	hemic and lymphatic diseases fungi Cell Biology Molecular Biology
Zdroj:	Molecular and Cellular Biology. 13:5469-5478
ISSN:	1098-5549 0270-7306
Popis:	The goal of this study was to determine whether it will be feasible to study the expression of a large, human gene, such as the BCL2 proto-oncogene, by DNA transfection. The BCL2 proto-oncogene is 230 kb in size and is deregulated in tumor cells by translocation into the immunoglobulin heavy-chain locus. Yeast artificial chromosomes (YACs) containing the human BCL2 gene were altered by homologous recombination in Saccharomyces cerevisiae to yield replicas of the normal and translocated alleles. Constructions containing either allele and ranging in size from 360 to 800 kb were integrated stably into a mouse tumor line. Fifty-eight percent of the clones contained a copy of the entire YAC insert. Over 50% of these clones expressed appropriate levels of human BCL2 RNA and protein. These studies suggested that the expression of large human genes and their pathologic rearrangements can be studied by transfection techniques employing YACs propagated in S. cerevisiae.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_________::705c54cf4ac57c67f1b9f1f512ac1072 https://doi.org/10.1128/mcb.13.9.5469 Zobrazit plný text záznamu