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Chicken hypersensitive site-4 insulator increases human serum albumin expression in bovine mammary epithelial cells modified with phiC31 integrase.
- Source :
-
Biotechnology letters [Biotechnol Lett] 2013 Apr; Vol. 35 (4), pp. 529-37. Date of Electronic Publication: 2012 Dec 22. - Publication Year :
- 2013
-
Abstract
- Achieving high expression levels of recombinant human serum albumin (HSA) for purification is a solution for the large amount of plasma-derived HSA needed in therapeutic applications. Here, we employed phiC31 integrase system and chicken hypersensitive site-4 (cHS4) insulators to construct a HSA expression vector for high-level HSA expression. The phiC31 integrase system mediated efficient transgene integration in bovine mammary epithelial cells (bMECs). A preferred pseudo attP site, which had 38 % identity with the 39 bp wild-type attP sequence, was detected in six out of 55 bMEC colonies. Addition of the cHS4 insulator to the phiC31 integrase system resulted in 8-20-fold increases of HSA expression compared with that of using integrase alone. Moreover, the reverse-oriented cHS4 insulator in the phiC31 integrase system provided the optimal level of HSA expression in bMECs.
- Subjects :
- Animals
Cattle
Cell Culture Techniques methods
Chickens
Humans
Insulator Elements
Integrases genetics
Integrases metabolism
Recombinant Proteins biosynthesis
Serum Albumin, Human
Biotechnology methods
Epithelial Cells metabolism
Gene Expression
Genetic Vectors
Serum Albumin biosynthesis
Technology, Pharmaceutical methods
Subjects
Details
- Language :
- English
- ISSN :
- 1573-6776
- Volume :
- 35
- Issue :
- 4
- Database :
- MEDLINE
- Journal :
- Biotechnology letters
- Publication Type :
- Academic Journal
- Accession number :
- 23264267
- Full Text :
- https://doi.org/10.1007/s10529-012-1125-y