Linked Life Data

7292260

Source:http://linkedlifedata.com/resource/pubmed/id/7292260

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
5
pubmed:dateCreated
1981-12-22
pubmed:abstractText
We have investigated several of the experimental factors that affect calcium phosphate-DNA-mediated gene transfer of thymidine kinase (tk) into mouse LM tk- Cl 1D cells using unfractionated DNA from both Chinese hamster ovary cells and L6 rat myoblasts. Increases in the length of exposure to DNA (24 h) and the expression time (48 h) before selection result in a 20-fold enhancement in the efficiency of transformation. These modifications yield frequencies up to 35 HATR colonies/20 microgram tk"NA/10(6) recipient cells. Exposure to dimethyl sulfoxide enhances transformation efficiencies slightly for short DNA exposure times, but has no effect when optimal DNA exposure times are used. Several other variations in our standard transformation protocol were also examined: these include the concentration and size of the DNA and exposure to low concentrations of the nonionic detergent, Tween-80. We have also isolated and characterized a subclone of Cl 1D that is a high-efficiency recipient for the tk+ marker. Segregation analysis reveals that the majority of the TK+ transformants derived from this subclone are stable, in contrast to those derived from the DL 1D parent. The combination of improved methodology and the high-efficiency recipient subclone permits DNA-mediated transformation for tk at frequencies on the order of 10(-4) transformants per recipient cell.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Sep
pubmed:issn
0098-0366
pubmed:author
pubmed:issnType
Print
pubmed:volume
7
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
603-16
pubmed:dateRevised
2007-11-14
pubmed:meshHeading
pubmed:year
1981
pubmed:articleTitle
Enhancing the efficiency of DNA-mediated gene transfer in mammalian cells.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S., Research Support, Non-U.S. Gov't