[Expression and purification of human apolipoprotein M].
OBJECTIVE
To express and purify the extra cellular full-length human apolipoprotein M(ApoM).


METHODS
The ApoM gene fragment was amplified from the human liver cDNA library by PCR. The resulting product was cloned into pGEXT vector and sequenced. Then the confirmed canstatin cDNA was cloned into plasmid E.coli JM109 and then transformed into E.coli DL21(DE3) where it was induced to express protein by IPTG.


RESULTS
The ApoM gene was cloned by PCR and a 560 bp DNA fragment was shown on the agarose electrophoresis. The cloned gene was sequenced and demonstrated to have the same sequence as that of human ApoM gene in GenBank. Then ApoM cDNA gene fragment was induced by IPTG, and a 24 kD recombinant ApoM protein was tested on SDS-PAGE.


CONCLUSIONS
Human ApoM gene is successfully cloned and its recombinant proteins are expressed.
Hu
Min
M
Department of Cardiovasology, Second Xiangya Hospital, Central South University, Changsha 410011,China.
Zhao
Shui-ping
SP
Zhang
Tao
T
Pan
Yi
Y
Xiong
Dan
D
D001053:Apolipoproteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification*; D000075243:Apolipoproteins M; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D054834:Lipocalins; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification
C400523:APOM protein, human; D001053:Apolipoproteins; D000075243:Apolipoproteins M; D018076:DNA, Complementary; D054834:Lipocalins; D011994:Recombinant Proteins