Application of mechano-growth factor in preparation of serum-free cultured tolerant type mammal engineering cell
A technology of serum-free culture and growth factors, applied in the field of cell engineering and genetic engineering recombination, to achieve the effect of reducing requirements, improving tolerance and broad application prospects
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[0017] In order to make the object, technical solution and advantages of the present invention clearer, preferred embodiments of the present invention will be described in detail below with reference to the accompanying drawings. Experimental methods not indicating specific conditions in the preferred embodiments are generally in accordance with conventional conditions, such as described in the Molecular Cloning Experiment Guide (Third Edition, J. Sambrook et al., translated by Huang Peitang, Science Press, 2002) conditions, or as recommended by the manufacturer.
[0018] 1. Construction and identification of engineered mammalian cells stably expressing mouse MGF
[0019] The mouse MGF coding gene whose nucleotide sequence is shown as SEQ ID No.4 was synthesized. Follow ViraPower TM Lentiviral Directional TOPO ? According to the instructions of Expression Kit (Invitrogen), the following primers were synthesized: upstream primer F: 5'-caccatgggaccggagacgc-3'; downstream ...
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