Development of an α-amylase reporter system for efficient screening of clones with highly expressed heterologous protein in Hansenula polymorpha.
To check feasibility and effectiveness of the α-amylase reporter system, two vectors were designed and tested using hepatitis B virus surface antigen (HBsAg) and Homo sapiens granulocyte-macrophage colony stimulating factor 2 (hGM-CSF2) as a model. By integrating the vector containing two independent cassettes into the same genome locus, high-producing clones of HBsAg (or hGM-CSF2) were screened using the α-amylase as a reporter. Results show there was a positive correlation (Correlation coefficient, R (2) > 0.95) between the yield of recombinant proteins and the α-amylase activity of corresponding transformants, which was independent of the gene dosage.
Song
Hao-Lei
HL
Institute of Microbiology, Chinese Academy of Sciences, No. 1 Beichen West Road, Chaoyang District, Beijing, 100101, China.
Niu
Zhen-Dong
ZD
Qian
Wei-Dong
WD
Wang
Hui
H
Qiu
Bing-Sheng
BS
D001709:Biotechnology / Q000379:methods*; D017930:Genes, Reporter; D005822:Genetic Vectors; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000096:biosynthesis / Q000235:genetics; D006514:Hepatitis B Surface Antigens / Q000096:biosynthesis / Q000235:genetics; D006801:Humans; D008403:Mass Screening / Q000379:methods; D010843:Pichia / Q000201:enzymology* / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D000516:alpha-Amylases / Q000096:biosynthesis* / Q000235:genetics
D006514:Hepatitis B Surface Antigens; D011994:Recombinant Proteins; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D000516:alpha-Amylases