[Construction of a retroviral vector carrying human growth hormone gene and its introduction and expression in 3T3 and ES cells].
In order to study the expression of growth hormone gene in heterologous cells, the retroviral vector pINS was used to construct the recombinant plasmid carrying the human growth hormone gene (hGH), and then it was introduced into mouse fibroblast cell line 3T3 and mouse embryonic stem cell line CCE. Levels of accumulation of hGH secreted into medium was quantified by a radioimmunoassay. Expression level of hGH gene varied considerably from one clone to another. Several 3T3 clones expressing hGH gene at high level were obtained, such as GHSNC 20, in which hGH concentration in medium was 3784ng/ml, Integration of exogenous gene was stable. Southern blot result showed that such kind of difference was not related to the number of integrated copies of the foreign gene. The expression of ES clones transfected with pINS-GH was undetectable, and integration of the foreign gene was unstable. Southern blot result showed that hGH gene really integrated into the cellular genome, and no rearrangement of foreign gene is found at present.
Zeng
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Y
Department of Biology, Peking University Beijing.
Wu
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H
D016475:3T3 Cells; D000818:Animals; D005822:Genetic Vectors; D013006:Growth Hormone / Q000096:biosynthesis / Q000235:genetics*; D006801:Humans; D051379:Mice; D012190:Retroviridae / Q000235:genetics; D013234:Stem Cells / Q000378:metabolism; D014162:Transfection
D013006:Growth Hormone