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Production and Characterization of Recombinant Mouse Brain‐Derived Neurotrophic Factor and Rat Neurotrophin‐3 Expressed in Insect Cells by Sheryl L. Meyer; Diane M. Lang; M. Elizabeth Forbes; Ernest Knight Jr.; James D. Hirsch; Stephen P. Trusko; Richard W. Scott is a Neuroscience article available to read on EtoBox.
What is Production and Characterization of Recombinant Mouse Brain‐Derived Neurotrophic Factor and Rat Neurotrophin‐3 Expressed in Insect Cells about?
Abstract: Bioactive brain‐derived neurotrophic factor (BDNF) and neurotrophin‐3 were produced using the baculovirus expression system and purified to homogeneity using ion‐exchange and reversed‐phase chromatography. Yields of purified neurotrophin‐3 (300–500 μg/L) were similar to levels reported for baculovirus‐expressed nerve growth factor (NGF), whereas initial yields of BDNF were significantly lower (20–50 μg/L). Improved production of BDNF (150–200 μg/L) was achieved by expressing BDNF from a chimeric prepro‐NGF/mature BDNF construct using the __Trichoplusia ni__ insect cell line, Tn‐5B1‐4. Examination of the distribution of BDNF protein from both the nonchimeric prepro‐BDNF and the chimeric prepro‐NGF/mature BDNF viruses in Sf‐21‐and Tn‐5B1‐4‐infected cells suggests a specific deficiency in the Tn‐5B1‐4 cells in processing the nonchimeric precursor. In addition, the vast majority of the BDNF protein at 2 days after infection was intracellular and insoluble. N‐terminal amino acid sequencing of purified recombinant BDNF and neurotrophin‐3 demonstrated that the insect cells processed their precursors to the correct N‐terminus expected for the mature protein. Bioactivity was char
Who reads Production and Characterization of Recombinant Mouse Brain‐Derived Neurotrophic Factor and Rat Neurotrophin‐3 Expressed in Insect Cells?
It is typically read by researchers, students, and practitioners in Neuroscience.
- Author
- Sheryl L. Meyer; Diane M. Lang; M. Elizabeth Forbes; Ernest Knight Jr.; James D. Hirsch; Stephen P. Trusko; Richard W. Scott
- Publisher
- John Wiley and Sons; Wiley (Blackwell Publishing); Blackwell Publishing Inc.; Wiley (ISSN 0022-3042)
- Published
- 1994
- Language
- EN
- Field
- Neuroscience (Life Sciences)