Cloning and characterization of 5'-untranslated region of porcine beta casein gene (CSN2).

Abstract:

:beta-Casein (CSN2) is a major milk protein in most mammals. The CSN2 gene is generally induced by lactogenic hormones bound to its promoter. The expression of this gene can be enhanced by signal transducers and activators of transcription (STAT) and glucocorticoid receptor (GR). Here, we analyzed the promoter and intron 1 regions of the porcine CSN2 gene. The porcine CSN2 promoter and intron 1 regions (-3098bp to +2446bp) were cloned into the pGL3-Basic vector containing the luciferase reporter gene (pCSN2-PEI). Lactogenic signals induced the transcription of porcine CSN2. By using AG490, a Janus kinase (JAK) inhibitor, we demonstrated that STAT5 positively regulates the transcription of porcine CSN2. Further, seven STAT mutants were generated by site-directed mutagenesis. By performing electrophoretic mobility shift assays (EMSAs), we located a critical element for pCSN2-PEI transcription bound to STAT5 in the -102bp to -84bp region. The construct containing only the promoter region (pCSN2-P), however, did not exert any promotive effects on transcription in two cell types-a mouse mammary epithelial cell line (HC11) and porcine mammary gland epithelial cells (PMECs). Thus, the construct containing intron 1 of porcine CSN2 exerts an elevating effect on transcription. We suggest that the transcription of porcine CSN2 is regulated by lactogenic signals via the STAT5 site (-102bp to -84bp) and intron 1.

journal_name

Domest Anim Endocrinol

authors

Lee P,Chung HK,Lee HG,Lee HC,Woo JS,Lee S,Jo SJ,Chang WK,Lee HT,Kwon M,Park JK

doi

10.1016/j.domaniend.2008.04.001

subject

Has Abstract

pub_date

2008-10-01 00:00:00

pages

245-53

issue

3

eissn

0739-7240

issn

1879-0054

pii

S0739-7240(08)00041-6

journal_volume

35

pub_type

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