[Expression of human leukemia inhibitory factor gene in Pichia pastoris].
The 575 bp artificial synthesized human leukemia inhibitory factor (hLIF) gene was cloned into vector pPICZ alpha A and the constructed expression vector pPICZ alpha A-hLIF was linearized by SacI and transformed into Pichia pastoris X-33. After the screening for Mut phenotype and the PCR analysis of the transformants, hLIF gene expressed in Pichia pastoris. SDS-PAGE and Western blot analysis of the culture supernatants showed that hLIF gene could express as 58.5 kD protein with the desired immunogenicity. Density scanning of the SDS-PAGE gel revealed that the targeted protein accounted for 32.8% of the total protein in the supernatants. The expressed products can inhibit the clone formation of murine's teratoma cells.
Wang
Q
Q
Biopharmaceutical Center, State Key Laboratory for Biological Control, Zhongshan University, Guangzhou 510275, China.
Yang
L
L
Quyang
J
J
Long
Q
Q
Wang
X
X
D000818:Animals; D003001:Cloning, Molecular; D004274:DNA, Recombinant / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors; D015850:Interleukin-6 / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology; D053605:Leukemia Inhibitory Factor; D051379:Mice; D010843:Pichia / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids; D011994:Recombinant Proteins / Q000235:genetics; D013724:Teratoma / Q000473:pathology; D014407:Tumor Cells, Cultured / Q000187:drug effects
D004274:DNA, Recombinant; D015850:Interleukin-6; C507837:LIF protein, human; D053605:Leukemia Inhibitory Factor; C507838:Lif protein, mouse; D011994:Recombinant Proteins