The tubulin beta III (
TUBB3) gene encodes a class III member of the beta tubulin protein family that is
primarily expressed in neurons and is considered to play a critical role in proper axon guidance and maintenance. This
protein is generally used as a specific marker of neurons in the central nervous system. We obtained the full length cDNA
sequence of
TUBB3 by using the RACE method based on the EST fragment from the brain and spinal cord cDNA library
of
Gekko japonicus
. We further investigated the multi-tissue expression pattern by RT-PCR and identified one transcript
of TUBB3 about 1.8 kb in the central nervous system of
Gekko japonicus by Northern blotting. The completed cDNA of
gecko TUBB3 is 1 790 bp with an open reading frame of 1 350 bp, encoding a 450 amino-acid protein. The recombinant
plasmid of pET-32a-TUBB3 was constructed and induced to express His-tagged TUBB3 protein in prokaryotic BL21
cells. The purified TUBB3 protein was then used to immunize rabbits to generate polyclonal antisera. The titer of the
antiserum was more than 1:65 536 determined by ELISA. The result of western blotting showed that the TUBB3 antibody
could specifically recognize the recombinant TUBB3 protein and endogenous TUBB3 protein. Our findings provide the tools
to further understand the
TUBB3 gene and investigate the regeneration of the central nervous system in
Gekko japonicas.
