Nucleic Acids Research, 1994, Vol. 22, No. 7 1281-1286
© 1994
MOLECULAR BIOLOGY |
Identification of a lens-specific regulatory region (LSR) of the murine 
B-crystallin gene
Laboratory of Molecular and Developmental Biology, National Eye Institute, National Institutes of Hearth Bethesda, MD 20892, USA
*To whom correspondence should be addressed
Received October 27, 1993. Revised March 4, 1994. Accepted March 4, 1994.
Previous studies have shown that the –661/ + 44 sequence of the murine 
B-crystallln gene contains a muscle-preferred enhancer (–426/–257) and can drive the bacterial chloramphenicol acetyltransferase (CAT) gene in the lens, skeletal muscle and heart of transgenlc mice. Here we show that transgenlc mice carrying a truncated -164/+ 44 fragment of the aB-crystallin gene fused to the CAT gene expressed exclusively in the lens; by contrast mice carrying a –426/+ 44 fragment of the aB gene fused to CAT expressed highly in the lens, skeletal muscle and heart, and slightly in the lung, brain, kidney, spleen and liver. DNase I protection experiments indicated that the –147–118 sequence is protected by nuclear proteins from TN4-1 lens cell line, but not by nuclear proteins from myotubes of the C2C12 cell line. Site directed mutagenesis of this sequence decreased promoter activity in translently-transfected lens cells, consistent with this sequence being a lens-specific regulatory region (LSR). We conclude that the –426–257 enhancer is required for expression in skeletal muscle, heart and possibly other tissues, and that the –164/+44 sequence of the B-crystallin gene is sufficient for expression in the lens of transgenic mice.
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