[Expression and purification of two kinds of alternative splicing mouse Era].
OBJECTIVE
To express and purify two alternative splicing mouse Era proteins and detect whether anti-human Era antibody can be used in the study of mouse Era proteins.


METHODS
Two fusion protein expression vectors, pMAL-meraW and pMAL-meraS, were constructed, then the MBP-mEra proteins were expressed in E. coli. The target proteins were purified by amylose affinity chromatography. The specificity of rabbit anti-human Era antibody to the proteins was identified by Western blot.


RESULTS
The expressed MBP-mEraW and MBP-mEraS proteins constituted approximately 17% and 19% of the total bacterial proteins. The purity of the fused proteins was 67% and 61% respectively after amylose affinity chromatography. Rabbit anti-human Era antibody had high specificity to these two kinds of splicing mouse Era proteins.


CONCLUSIONS
Two fusion mera genes could be expressed in E. coli by using gene recombination technique. The high specificity of rabbit anti-human Era antibody to the two splicing mouse ERA proteins indicates that this antibody can be used to study the function of these two kinds of splicing mouse Era.
Dong
Ke
K
Department of Biochemistry and Molecular Biology, Fourth Military Medical University, Xi'an 710032, China.
Chen
Su-min
SM
Ji
Zong-ling
ZL
Zheng
Yu
Y
Chen
Nan-chun
NC
D017398:Alternative Splicing; D000818:Animals; D015153:Blotting, Western; D004587:Electrophoresis, Agar Gel; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D019204:GTP-Binding Proteins; D015870:Gene Expression; D005818:Genetic Engineering; D006801:Humans; D051379:Mice; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D016601:RNA-Binding Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification
C540137:Era protein, mouse; D016601:RNA-Binding Proteins; D011993:Recombinant Fusion Proteins; D019204:GTP-Binding Proteins