Sequential cloning of chromosomes Metadata
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- Main Title Sequential cloning of chromosomes
Inventor: Lacks, S.A.Creator Type: Personal
Assignee: United States. Department of Energy.Contributor Type: OrganizationContributor Info: Patent Assignee: Dept. of Energy; USDOE, Washington, DC (United States)
Name: Brookhaven National LaboratoryPlace of Publication: Upton, New YorkAdditional Info: Brookhaven National Lab., Upton, NY (United States)
- Creation: 1991-12-31
- Content Description: A method for sequential cloning of chromosomal DNA and chromosomal DNA cloned by this method are disclosed. The method includes the selection of a target organism having a segment of chromosomal DNA to be sequentially cloned. A first DNA segment, having a first restriction enzyme site on either side. homologous to the chromosomal DNA to be sequentially cloned is isolated. A first vector product is formed by ligating the homologous segment into a suitably designed vector. The first vector product is circularly integrated into the target organism`s chromosomal DNA. The resulting integrated chromosomal DNA segment includes the homologous DNA segment at either end of the integrated vector segment. The integrated chromosomal DNA is cleaved with a second restriction enzyme and ligated to form a vector-containing plasmid, which is replicated in a host organism. The replicated plasmid is then cleaved with the first restriction enzyme. Next, a DNA segment containing the vector and a segment of DNA homologous to a distal portion of the previously isolated DNA segment is isolated. This segment is then ligated to form a plasmid which is replicated within a suitable host. This plasmid is then circularly integrated into the target chromosomal DNA. The chromosomal DNA containing the circularly integrated vector is treated with a third, retrorestriction enzyme. The cleaved DNA is ligated to give a plasmid that is used to transform a host permissive for replication of its vector. The sequential cloning process continues by repeated cycles of circular integration and excision. The excision is carried out alternately with the second and third enzymes.
- Physical Description: 42 p.
- Keyword: Plasmids
- Keyword: Genetic Mapping
- Keyword: Dna Sequencing
- Keyword: Dna-Cloning
- STI Subject Categories: 55 Biology And Medicine, Basic Studies
- Other Information: PBD: 1991
Name: Office of Scientific & Technical Information Technical ReportsCode: OSTI
Name: UNT Libraries Government Documents DepartmentCode: UNTGD
- Other: DE94007362
- Report No.: PATENTS-US--A7811221
- Grant Number: AC02-76CH00016
- Office of Scientific & Technical Information Report Number: 140887
- Archival Resource Key: ark:/67531/metadc624275
- Display Note: OSTI as DE94007362