Development of an [alpha]-amylase reporter system for efficient screening of clones with highly expressed heterologous protein in Hansenula polymorpha

To check feasibility and effectiveness of the a-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 independen...

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Bibliographic Details
Published inBiotechnology letters Vol. 32; no. 10; pp. 1473 - 1479
Main Authors Song, Hao-Lei, Niu, Zhen-Dong, Qian, Wei-Dong, Wang, Hui, Qiu, Bing-Sheng
Format Journal Article
LanguageEnglish
Published Dordrecht Springer Nature B.V 01.10.2010
Subjects
Antigens
Biotechnology
Cloning
Correlation coefficient
Genes
Genomes
Hansenula polymorpha
Hepatitis
Hepatitis B virus
Mathematical analysis
Proteins
Recombinant
Vectors (mathematics)
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Summary:To check feasibility and effectiveness of the a-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 a-amylase as a reporter. Results show there was a positive correlation (Correlation coefficient, R super(2)>0.95) between the yield of recombinant proteins and the a-amylase activity of corresponding transformants, which was independent of the gene dosage.
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ISSN:0141-5492
1573-6776
DOI:10.1007/s10529-010-0314-9