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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
20
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pubmed:dateCreated |
1995-10-27
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pubmed:abstractText |
Expression cloning of cDNAs was first described a decade ago and was based on transient expression of cDNA libraries in COS cells. In contrast to transient transfection of plasmids, retroviral gene transfer delivers genes stably into a wide range of target cells. We utilize a simple packaging system for production of high-titer retrovirus stock from cDNA libraries to establish a cDNA expression cloning system. In two model experiments, murine interleukin (IL)-3-dependent Ba/F3 cells were infected with libraries of retrovirally expressed cDNA derived from human T-cell mRNA or human IL-3-dependent TF-1 cell line mRNA. These infected Ba/F3 cells were selected for the expression of CD2 by flow cytometry or for the alpha subunit of the human IL-3 receptor (hIL-3R alpha) by factor-dependent growth. CD2 (frequency, 1 in 10(4)) and hIL-3R alpha (frequency, 1 in 1.5 x 10(5)) cDNAs were readily detected in small-scale experiments, indicating this retroviral expression cloning system is efficient enough to clone low-abundance cDNAs by their expression or function.
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pubmed:grant |
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/7568090-1377056,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7568090-1408844,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7568090-1833064,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7568090-2158861,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7568090-2194165,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7568090-2404337,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7568090-2437578,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7568090-2539263,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7568090-2598259,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7568090-2663885,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7568090-2725678,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7568090-3136546,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7568090-3457477,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7568090-3479791,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7568090-3918306,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7568090-6260373,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7568090-6300662,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7568090-6331674,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7568090-7690960,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7568090-7823939,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7568090-8057434,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7568090-8289827
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
Sep
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pubmed:issn |
0027-8424
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:day |
26
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pubmed:volume |
92
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
9146-50
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pubmed:dateRevised |
2009-11-18
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pubmed:meshHeading |
pubmed-meshheading:7568090-3T3 Cells,
pubmed-meshheading:7568090-Animals,
pubmed-meshheading:7568090-Base Sequence,
pubmed-meshheading:7568090-Cell Line,
pubmed-meshheading:7568090-Cercopithecus aethiops,
pubmed-meshheading:7568090-Cloning, Molecular,
pubmed-meshheading:7568090-DNA, Complementary,
pubmed-meshheading:7568090-Databases, Factual,
pubmed-meshheading:7568090-Efficiency,
pubmed-meshheading:7568090-Genetic Vectors,
pubmed-meshheading:7568090-Humans,
pubmed-meshheading:7568090-Interleukin-3,
pubmed-meshheading:7568090-Kidney,
pubmed-meshheading:7568090-Mice,
pubmed-meshheading:7568090-Molecular Sequence Data,
pubmed-meshheading:7568090-Oligodeoxyribonucleotides,
pubmed-meshheading:7568090-RNA, Messenger,
pubmed-meshheading:7568090-Receptors, Interleukin-3,
pubmed-meshheading:7568090-Recombinant Proteins,
pubmed-meshheading:7568090-Restriction Mapping,
pubmed-meshheading:7568090-Retroviridae,
pubmed-meshheading:7568090-T-Lymphocytes,
pubmed-meshheading:7568090-Transfection
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pubmed:year |
1995
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pubmed:articleTitle |
Efficient screening of retroviral cDNA expression libraries.
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pubmed:affiliation |
Department of Cell Biology, DNAX Research Institute of Molecular and Cell Biology, Palo Alto, CA 94304, USA.
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.,
Research Support, Non-U.S. Gov't
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