[High efficient expression of recombinant human platelet factor IV in Pichia pastoris].
OBJECTIVE
To express recombinant human platelet factor 4 (PF4) in methanol yeast.


METHODS
PF4 cDNA amplified by PCR was cloned into the mating factor alpha (MFalpha) signal sequence, downstream the alcohol oxidase 1 (AOX 1) promoter of the pPIC9 vector. The reconstructed vector was subsequently integrated into the methanol yeast P. pastoris strain GS115. The highly expressing strain was selected and the protein sequences and their biological activities were assayed.


RESULTS
The amino acid sequence of rhPF4 was the same as that of natural one. The rhPF4 can neutralize the anticoagulation effect of heparin in a dose-dependent manner.


CONCLUSIONS
The amount of rhPF4 expressed in methanol yeast meets well the scale for manufacturing. The biological activities of the rhPF4 were the same as that of natural PF4.
Wang
C
C
Research Department of Jiuyuan Gene Engineering Co. LTD, Hangzhou 310018, China.
Xie
Y
Y
Wang
T
T
Yang
W
W
Li
C
C
Zhou
J
J
D003001:Cloning, Molecular; D015870:Gene Expression; D006801:Humans; D010843:Pichia / Q000235:genetics* / Q000378:metabolism; D010978:Platelet Factor 4 / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism
D011994:Recombinant Proteins; D010978:Platelet Factor 4