PURIFIED RECOMBINANT GLIAL CELL-DERIVED NEUROTROPIC FACTOR (GDNF) OF ROHU CARP, LABEO ROHITA

摘要

Mammalian glial cell line-derived neurotrophic factor (GDNF) has been reported to promote stem cell proliferation. Recombinant GDNF from rat, mouse and human origins are available commercially. These rGDNFs fail to promote proliferation of rohu spermatogonial stem cells. GDNF from teleost are not available commercially. In this study, recombinant rohu glial cell line-derived neurotrophic factor (RrGDNF) has been expressed at high levels and produced in large quantities in Baculovirus infected Sf21 insect cells. The expressed recombinant protein (His-tagged) was purified using proBond resin in a denaturing condition. The protein was stabilized with the help of dialyzing in a PBS buffer. The purified protein was confirmed by SDS-PAGE and Western blotting using anti-His antibody. Pure, recombinant GDNF promoted the colonization and proliferation of rohu spermatogonial stem cells in vitro. To the best of our knowledge, this is the first recombinant GDNF protein of fish species that can be utilized for basic biology research. The purified RrGDNF (rohu recombinant GDNF) protein can be obtained in large quantities having commercial implications.