The human kiaa0319-like gene is one of the candidate genes for developmental dyslexia, but the exact function of the encoded KIAA0319L (KL) protein is not known. To allow functional analysis a purified, biologically active KL protein is required. The kiaa0319-like gene was expressed in insect cells using the baculovirus expression system. To optimize the expression of the kiaa0319-like gene and to be able to purify the KL protein, several approaches were used. Two different recombinant baculoviruses were made, one with the full length coding sequence of KL and one that lacked the transmembrane domain to facilitate purification.. Versions in which the kiaa0319L sequences were cloned downstream of the honeybee melittin signal sequence were also made. All four constructs contained a C-terminal influenza hemagglutinin (HA)-tag. Sf9 insect cells infected with these recombinant baculoviruses produced the KL protein, as demonstrated by Western blot analysis using either the HA-antibody or KL-specific polyclonal serum.
|Journal||Journal of Biochemistry and Molecular Biology in The Post Genomic Era|
|Publication status||Published - 2013|