[Expression of a candidate human Era binding protein A19 and the preparation of its polyclonal antibody].
OBJECTIVE
To express a candidate hEra binding protein A19 in Escherichia coli and to prepare anti-A19 antibody.


METHODS
A19 gene was amplified by PCR from the plasmid containing A19 gene and was cloned into the expression vector pGEX-4T3 which was then transformed into E.coli. The A19 protein was expressed under IPTG induction. Antiserum was prepared by immunizing rabbits with the expressed A19 protein. The titer and specificity of polyclonal antibody were detected by Western blot.


RESULTS
The expressed A19 accounted for about 30.2% of total bacterial protein. The titer of the antiserum was about 1:4 000. Western blot analysis indicated that the antiserum had high specificity.


CONCLUSIONS
A19 fusion protein was highly expressed. The specific anti-A19 antiserum was prepared successfully.
Department of Biochemistry and Molecular Biology, Fourth Military Medical University, Xi'an 710032, China. Liyf@263.net
Yun-feng
YF
Li
Yuan-ming
YM
Wu
Su-min
SM
Chen
Nan-chun
NC
Chen
Yong
Y
Huang
Xiao-nan
XN
Zhang
D000818:Animals; D000918:Antibody Specificity; D015153:Blotting, Western; D002352:Carrier Proteins / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism; D004262:DNA Restriction Enzymes / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D006801:Humans; D007106:Immune Sera / Q000032:analysis / Q000276:immunology*; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D016133:Polymerase Chain Reaction; D016601:RNA-Binding Proteins / Q000378:metabolism*; D020798:Two-Hybrid System Techniques
C540138:A19 protein, human; D002352:Carrier Proteins; D007106:Immune Sera; D016601:RNA-Binding Proteins; D004262:DNA Restriction Enzymes