Tissue-specific expression of human salivary mucin gene, MUC7, in transgenic mice.
The MUC7 gene encodes the protein core of the low molecular weight human salivary mucin (MG2, mucin glycoprotein 2) and is expressed in a tissue-specific manner in salivary glands. The purpose of this study was to examine MUC7 expression by transgenic mouse technology. A 16 kb DNA fragment, containing the MUC7 gene (10 kb) and 3 kb of the upstream and 3 kb of the downstream sequences, was used to generate transgenic mice. We have identified five transgenic founder mice which were propagated as individual transgenic lines and analysed. Tissues of transgenic offspring from each line were analysed by RT-PCR to determine the sites of the MUC7 expression. The results indicated that only line 3 and line 5 expressed the MUC7 gene in salivary glands. The level of MUC7 expression in selected tissues was then determined by northern blot analyses. The results showed that line 3 mice contained high levels of MUC7 transcripts in the sublingual glands of both males and females and indicated low levels of MUC7 transcripts in the submandibular glands of females. No MUC7 expression was detected in this line by northern blot analysis in any other tissue tested. On the other hand, no expression of MUC7 was detected in any tissues of line 5 mice examined by northern blot analysis. A Southern blot analysis of human and mouse genomic DNA demonstrated multiple copies of the MUC7 transgene in line 3 and a single copy in line 5. Collectively, these results indicate that the regulatory sequences required for the tissue-specific expression of MUC7 are within the MUC7 transgene. However, the sequences necessary for expression comparable to that of MUC7 in human salivary glands may be missing from this construct. Western blot analysis of protein extracts from different tissues of transgenic mice line 3 showed that MUC7 gene product was produced in the submandibular-sublingual gland complex of both male and female mice and not in the other tissues examined.
Bobek
L A
LA
Department of Oral Biology, State University of New York at Buffalo 14214, USA.
Li
H
H
Rojstaczer
N
N
Jones
C
C
Gross
K W
KW
Levine
M J
MJ
D000818:Animals; D015152:Blotting, Northern; D015139:Blotting, Southern; D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005786:Gene Expression Regulation; D006801:Humans; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D009077:Mucins / Q000096:biosynthesis / Q000235:genetics*; D009928:Organ Specificity; D016133:Polymerase Chain Reaction / Q000379:methods; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012471:Salivary Proteins and Peptides / Q000096:biosynthesis / Q000235:genetics*; D013361:Sublingual Gland / Q000378:metabolism*; D013363:Submandibular Gland / Q000378:metabolism*; D019076:Transgenes
C083238:MUC7 protein, human; D009077:Mucins; D012471:Salivary Proteins and Peptides