|
rdf:type |
|
|
lifeskim:mentions |
|
|
pubmed:issue |
1
|
|
pubmed:dateCreated |
1990-11-20
|
|
pubmed:abstractText |
Although many different protocols for transfection of lymphoid cell lines exist, successful DNA transfer into primary B lymphocytes has, to date, not been demonstrated. We now describe a simple method for gene transfer into highly purified normal and malignant B lymphocytes by electroporation. Using a powerful expression vector containing two copies of the cytomegalovirus (CMV) immediate early enhancer linked to the human T cell lymphotropic virus I (HTLV I) promoter, we could demonstrate transfected gene expression in both high density small 'resting' B cells and in low density 'activated' B cells. Successful transfection was detected by expression of chloramphenicol acetyl transferase and by immunofluorescence. The neoplastic cells of B cell chronic lymphocytic leukemia could also be transfected with an efficiency of 5-10%, but only after preactivation. This method of transfection will permit analysis of the contribution of individual genes and their products to normal and malignant B cell growth and differentiation.
|
|
pubmed:grant |
|
|
pubmed:language |
eng
|
|
pubmed:journal |
|
|
pubmed:citationSubset |
IM
|
|
pubmed:chemical |
|
|
pubmed:status |
MEDLINE
|
|
pubmed:month |
Oct
|
|
pubmed:issn |
0022-1759
|
|
pubmed:author |
|
|
pubmed:issnType |
Print
|
|
pubmed:day |
4
|
|
pubmed:volume |
133
|
|
pubmed:owner |
NLM
|
|
pubmed:authorsComplete |
Y
|
|
pubmed:pagination |
77-85
|
|
pubmed:dateRevised |
2009-9-29
|
|
pubmed:meshHeading |
pubmed-meshheading:1698880-Antigens, CD5,
pubmed-meshheading:1698880-Antigens, Differentiation,
pubmed-meshheading:1698880-B-Lymphocytes,
pubmed-meshheading:1698880-Cell Separation,
pubmed-meshheading:1698880-Chloramphenicol O-Acetyltransferase,
pubmed-meshheading:1698880-Cytomegalovirus,
pubmed-meshheading:1698880-Electric Stimulation,
pubmed-meshheading:1698880-Enhancer Elements, Genetic,
pubmed-meshheading:1698880-Fluorescent Antibody Technique,
pubmed-meshheading:1698880-Gene Expression,
pubmed-meshheading:1698880-Herpesvirus 4, Human,
pubmed-meshheading:1698880-Human T-lymphotropic virus 1,
pubmed-meshheading:1698880-Humans,
pubmed-meshheading:1698880-Leukemia, Lymphocytic, Chronic, B-Cell,
pubmed-meshheading:1698880-Lymphocyte Activation,
pubmed-meshheading:1698880-Plasmids,
pubmed-meshheading:1698880-Tetradecanoylphorbol Acetate,
pubmed-meshheading:1698880-Transfection,
pubmed-meshheading:1698880-Tumor Cells, Cultured
|
|
pubmed:year |
1990
|
|
pubmed:articleTitle |
Transfection and gene expression in normal and malignant primary B lymphocytes.
|
|
pubmed:affiliation |
Ludwig Institute for Cancer Research, St. Mary's Branch, London, U.K.
|
|
pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.,
Research Support, Non-U.S. Gov't
|
