[High expression of human thrombopoietin mutant fusion protein in E. coli].
OBJECTIVE
To obtain human thrombopoietin (Tpo)mutant cDNA encoding mature peptide N terminal 1 approximately 196 amino acids and investigate its expression in E. coli JM109.


METHODS
Polymerase chain reaction and DNA recombination techniques were employed. PCR product was inserted into pUC19 vector and sequenced and then cloned into expression vector pMAL-c2.


RESULTS
E. coli JM109 cells with plasmid pMAL -MBP/TpoM were induced by IPTG for 4-5 hours. SDS-PAGE analysis showed that MBP/TpoM molecular weight is about 63KD. Scanning analysis indicated that expressed protein accounts up to 37% of total E. coli proteins.


CONCLUSIONS
The Tpo mutant of interest is successfully expressed in E. coli JM109 cells.
Tian
S
S
Norman Bethune University of Medical Sciences, Changchun 130021.
Liu
L
L
Feng
B
B
D018076:DNA, Complementary / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D006801:Humans; D009154:Mutation; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D013926:Thrombopoietin / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification
D018076:DNA, Complementary; D011993:Recombinant Fusion Proteins; D013926:Thrombopoietin