Development of a set of expression vectors in Hansenula polymorpha.
Four expression vectors based on formate dehydrogenase promoter (FMDp) and methanol oxidase promoter (MOXp) from Hansenula polymorpha were developed to express heterologous genes in Hansenula polymorpha. A secretion signal sequence of the mating factor-alpha from Saccharomyces cerevisiae was inserted in the secretory expression plasmids for efficient secretion. A modified green fluorescent protein (mGFP5) was used as the marker of expression for the first time in H. polymorpha NCYC495 (leu 1.1) to determine the expression ability of these plasmids. The mGFP5 thus expressed retained its biochemical and physiological properties, such as accumulation inside cells and efficient secretion into the culture media. These results indicated that the four integrative vectors are useful expression systems which could be directly applied for production of heterologous proteins of interests in H. polymorpha.
Song
Houhui
H
Molecular Microbiological Centre, Institute of Microbiology, Chinese Academy of Sciences, 13# Zhongguancun Beiyitiao, Beijing 100080, PR China.
Li
Yong
Y
Fang
Weihuan
W
Geng
Yunfeng
Y
Wang
Xu
X
Wang
Min
M
Qiu
Bingsheng
B
D000429:Alcohol Oxidoreductases / Q000235:genetics* / Q000378:metabolism*; D003001:Cloning, Molecular / Q000379:methods*; D005560:Formate Dehydrogenases / Q000235:genetics* / Q000378:metabolism*; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology; D015966:Gene Expression Regulation, Fungal / Q000502:physiology; D005822:Genetic Vectors; D010843:Pichia / Q000201:enzymology* / Q000235:genetics*; D011401:Promoter Regions, Genetic; D015202:Protein Engineering / Q000379:methods; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics
D011993:Recombinant Fusion Proteins; D000429:Alcohol Oxidoreductases; C029293:methanol oxidase; D005560:Formate Dehydrogenases