Production of feline leukemia inhibitory factor with biological activity in Escherichia coli

Leukemia inhibitory factor (LIF) is a cytokine which is essential for oocyte and embryo development, embryonic stem cell, and induced pluripotent stem cell maintenance. Leukemia inhibitory factor improves the maturation of oocytes in the human and the mouse. However, feline LIF (fLIF) cloning and ef...

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Bibliographic Details
Published in:Theriogenology 2016-07, Vol.86 (2), p.604-611
Main Authors: Kanegi, R., Hatoya, S., Tsujimoto, Y., Takenaka, S., Nishimura, T., Wijewardana, V., Sugiura, K., Takahashi, M., Kawate, N., Tamada, H., Inaba, T.
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Animals
Base Sequence
Cats - physiology
Cloning, Molecular
Cumulus expansion
DNA, Complementary
Embryo, Mammalian - cytology
Escherichia coli - metabolism
Feline
Fibroblasts - physiology
Gene Expression Regulation, Bacterial - physiology
IVM
Leukemia inhibitory factor
Leukemia Inhibitory Factor - genetics
Leukemia Inhibitory Factor - metabolism
Nuclear maturation
Plasmids
Recombinant protein
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Summary:Leukemia inhibitory factor (LIF) is a cytokine which is essential for oocyte and embryo development, embryonic stem cell, and induced pluripotent stem cell maintenance. Leukemia inhibitory factor improves the maturation of oocytes in the human and the mouse. However, feline LIF (fLIF) cloning and effects on oocytes during IVM have not been reported. Thus, we cloned complete cDNA of fLIF and examined its biological activity and effects on oocytes during IVM in the domestic cat. The aminoacid sequence of fLIF revealed a homology of 81% or 92% with that of mouse or human. The fLIF produced by pCold TF DNA in Escherichia coli was readily soluble and after purification showed bioactivity in maintaining the undifferentiated state of mouse embryonic stem cells and enhancing the proliferation of human erythrocyte leukemia cells. Furthermore, 10- and 100-ng/mL fLIF induced cumulus expansion with or without FSH and EGF (P 
ISSN:0093-691X
1879-3231
DOI:10.1016/j.theriogenology.2016.02.013