The invention discloses a method of the in-vitro expression of a gekko japonicus Hoxc10 (homebox
gene c10)
protein and the preparation of a polyclonal
antibody, comprising the following steps of: obtaining an EST (
Expressed Sequence Tag) sequence of gekko japonicus Hoxc10; obtaining an overall length sequence of the gekko japonicus Hoxc10 through an RACE (
rapid amplification of cDNA ends) method; predicting an antigenic
epitope of the gekko japonicus Hoxc10
protein on line by adopting DNASTAR
software and SWISS-PLOT; constructing a
prokaryotic expression vector of the gekko japonicus Hoxc10 by selecting superior
epitope peptides; inducing the expression of fusion proteins
in vitro; purifying the fusion proteins; preparing the polyclonal
antibody, and the like. In the invention, an in-vitro gekko japonicus Hoxc10 expression
system constructed on the basis of pGEX-4T1 can efficiently express a
target protein in BL21 and further purify the
target protein; and a large quantity of GST-Hoxc10 fusion proteins can be conveniently obtained; in addition, a rabbit gekko japonicus Hoxc10
antiserum prepared by immunizing a
New Zealand rabbit through the GST-Hoxc10 fusion proteins has
high titer and good specificity.