Restriction map of a 35-kb HLA fragment constructed by nested deletion 'drop-out' mapping.
Gene
; 164(2): 335-9, 1995 Oct 27.
Article
em En
| MEDLINE
| ID: mdl-7590353
ABSTRACT
An efficient method for generating detailed restriction maps of large cloned DNA segments is demonstrated. The mapping strategy entails comparing restriction fragments from a parent clone and from nested deletion derivatives of that clone. In a set of deletion plasmids of decreasing size, an individual fragment will be lost, or 'drop-out', according to its position in the cloned fragment. In this demonstration, nested deletions were generated in both directions in a 35-kb DNA segment from the human leukocyte antigen (HLA) region by intramolecular transposition of an engineered gamma delta (Tn1000) element present in a special 'deletion factory' cloning vector [Wang et al., Proc. Natl. Acad. Sci. USA 90 (1993) 7874-7878]. Fifteen plasmids with deletions extending in one direction and eleven plasmids with deletions extending in the opposite direction were digested singly by each of four restriction enzymes. A total of 36 cleavage sites were mapped in the 35-kb HLA fragment. This drop-out approach using nested deletions provides a simple and efficient means of mapping restriction sites, genes and other features of interest in cosmid-sized cloned DNA segments or DNAs.
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Base de dados:
MEDLINE
Assunto principal:
Hominidae
/
Mapeamento por Restrição
/
Antígenos HLA
/
Complexo Principal de Histocompatibilidade
Limite:
Animals
/
Humans
Idioma:
En
Ano de publicação:
1995
Tipo de documento:
Article