Secretory expression of heterologous protein in Kluyveromyces cicerisporus.
To explore the potential of heterologous protein expression in Kluyveromyces cicerisporus, three expression plasmids, pUK1-PIT, pUKD-PIT and pUKD-S-PIT, based on the vector pUK1 or pUKD were constructed and transformed, respectively, into yeast strain K. cicerisporus Y179U. Human interferon alpha-2a, used as an example protein, was successfully expressed and secreted by transformant Y179U/pUKD-PIT and Y179U/pUKD-S-PIT. In the flask culture, strain Y179U/pUKD-S-PIT could express interferon at 60 mg/l. The stability of plasmid pUKD-S-PIT in the host was higher than that of pUKD-PIT. This was consistent with their expression levels of interferon. There were two interferon-related bands found by Western blotting analysis. The possible reason for this is discussed.
Cai
X P
XP
Institute of Genetics, State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University, Shanghai 200433, China.
Zhang
J
J
Yuan
H-Y
HY
Fang
Z-A
ZA
Li
Y-Y
YY
D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D000077190:Interferon alpha-2; D016898:Interferon-alpha / Q000032:analysis / Q000096:biosynthesis* / Q000235:genetics*; D007716:Kluyveromyces / Q000235:genetics* / Q000378:metabolism*; D010957:Plasmids; D015202:Protein Engineering; D011994:Recombinant Proteins
D000077190:Interferon alpha-2; D016898:Interferon-alpha; D011994:Recombinant Proteins