Construction of Mammary Gland-specific and Effective Expression Vector for Mammary Gland Bioreactor

doi:10.13523/j.cb.20140708

China Biotechnology

2014, Vol. 34

Issue (7): 49-55 DOI: 10.13523/j.cb.20140708

Construction of Mammary Gland-specific and Effective Expression Vector for Mammary Gland Bioreactor

ZHANG Si-min¹, GAO Yue¹, FANG Yu-dan^1,2, ZHANG Jin-mai¹, ZHANG Jin-zhi^1,2

1. Shanghai Children's Hospital, Shanghai Jiao Tong University, Shanghai Institute of Medical Genetics, Shanghai Jiao Tong University, Shanghai 200040, China;
2. Key Laboratory of Embryo Molecular Biology, Ministry of Health & Shanghai Key Laboratory of Embryo and Reproduction Engineering, Shanghai 200040, China

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Abstract

Mammary gland bioreactor is considered as a promising approach for generating pharmaceutical proteins. Yet, the low-level expression of the transgene is one of the bottlenecks in this technique. Here, a strong and constitutively expressing promoter instead of a mammary gland specific but weak promoter was used to improve transgene expression; meanwhile, a Cre/loxP system driven by a β-lactoglobulin (BLG) gene promoter was used to restrict the strong expression in mammary gland. Methods: Construct the mammary gland-specific expression unit (Polyadenylation signal-pBLG-cre), this unit was then flanked by a pCAG strong promoter and a lacZ reporter. Thus, a mammary gland-restricted expression vector named pCBCZ (pCAG-loxP-PolyA-pBLG-cre-loxP-lacZ) was constructed. The in vitro experiment results: Cre-loxP recombination was detected in the mouse epithelial (HC11) cells transfected with pCBCZ vector. X-Gal staining experiment showed that the vector can strongly drive β-galactosidase expression in HC11 cells, but weak in NIH 3T3 cells. These data indicate that the pCBCZ vector can effectively drive gene expression at a mammary gland-restricted manner, which provide a new strategy for the generation of an effective mammary gland bioreactor.

Key words： Mammary gland bioreactor Cre/loxP system Tissue specific expression lacZ reporter

Received: 21 February 2014 Published: 25 July 2014

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Cite this article:

ZHANG Si-min, GAO Yue, FANG Yu-dan, ZHANG Jin-mai, ZHANG Jin-zhi. Construction of Mammary Gland-specific and Effective Expression Vector for Mammary Gland Bioreactor. China Biotechnology, 2014, 34(7): 49-55.

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https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20140708 OR https://manu60.magtech.com.cn/biotech/Y2014/V34/I7/49

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