ISSN:
1432-0614
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
,
Process Engineering, Biotechnology, Nutrition Technology
Notes:
Abstract In order to obtain high-level expression of recombinant human neurotrophin-3 (NT-3), we constructed several types of expression plasmids and examined several cell lines for expression of the human NT-3 gene. The highest level production of the recombinant protein was attained in Chinese hamster ovary cells transfected with an expression plasmid that contains a chimera gene encoding the human nerve growth factor (NGF) prepro-region and human NT-3 mature-region under control of a murine leukemia virus-derived long terminal repeat (MuLV-LTR). This cell line can produce more than 1 mg recombinant human NT-3/1 conditioned medium. The recombinant protein was purified to apparent homogeneity with a cation exchange column, a gel filtration column and a reversed-phase HPLC column with a recovery of about 30%. The purified NT-3, at a concentration as low as 0.2 ng/ml, induced neurite out-growth in neurons prepared from 8-day-old chick embryonic dorsal root ganglia; however, it showed little neurotrophic effect on rat PC12 pheochromocytoma cells, which are known to be NGF-responding cells. In addition, this protein promoted colony formation by human peripheral blood lymphocytes in soft agar culture.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF00186964
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