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Analysis of Transgenic Mice Containing the Cystatin S Promoter
Author(s) -
Shaw Phyllis A.,
Li Jinhua,
Luo Jian
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.1_supplement.983.5
Subject(s) - messenger rna , biology , microbiology and biotechnology , sublingual gland , transgene , salivary gland , promoter , gene , gene expression , spleen , green fluorescent protein , submandibular gland , endocrinology , immunology , genetics , biochemistry
Phylogenetic “footprinting” of the cystatin S gene suggests that DNA‐binding sequences for specific functions are clustered in the 5′‐flanking region. The 1.9kb promoter has 3 types of transcription factor binding “modules”, including a hormone module containing three regions found in the 5′‐flanking sequence of all known salivary gland specific genes. To determine which of the cis ‐elements are functional and responsible for tissue specific regulation of the cystatin S gene, we generated two transgenic mouse models. One, the 1.9kb/GFP expressing transgene, contains the hormone module and the salivary gland‐specific elements. We examined the 1.9kb promoter driven expression of GFP mRNA during development of the submandibular, sublingual, and parotid glands, and liver, heart, kidney, muscle, spleen, brain, and lung, using real time RT‐PCR analysis of total RNA. The cystatin S 1.9kb promoter regulates GFP mRNA expression in all the tissues examined during development, albeit at a low, perhaps basal, level in some. Submandibular and sublingual glands exhibit the highest levels of GFP mRNA at 10, 15 and 25 days of development. The spleen, heart, muscle and lung have the highest level of GFP mRNA at day 10. The brain shows the highest level of GFP mRNA at 25 days; the parotid glands, liver and kidney show the highest level of GFP mRNA at 35 days. We are currently examining GFP protein levels in the same tissues during development of the 1.9kb/GFP transgenic mice to determine if tissue‐ and cell type‐specific regulation is at the protein level rather than the mRNA level.