TISSUE-SPECIFIC EXPRESSION AND METHYLATION OF A THYROGLOBULIN-CHLORAMPHENICOL ACETYLTRANSFERASE FUSION GENE IN TRANSGENIC MICE

被引:51
作者
LEDENT, C [1 ]
PARMENTIER, M [1 ]
VASSART, G [1 ]
机构
[1] UNIV LIBRE BRUXELLES,SERV GENET MED,B-1070 BRUSSELS,BELGIUM
关键词
Chloramphenicol acetyltransferase assay; DNA methylation; Thyroglobulin gene; Transcription control cAMP;
D O I
10.1073/pnas.87.16.6176
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Fusion genes containing 1600 or 2000 base pairs of the bovine thyroglobulin gene 5′ flanking region and the chloramphenicol acetyltransferase (CAT) coding sequence were constructed and used to generate transgenic mice. Altogether, 24 independent transgenic lines were obtained, and the expression of the transgene was assayed by measuring the CAT activity in different tissues. Depending on the transgenic lines, the fusion gene was either silent in all tissues or specifically expressed in the thyroid. The level of expression was found to be highly variable from one line to another and to be regulated by thyrotropin in a manner similar to the natural thyroglobulin gene. The methylation status of the integrated DNA was tested by digestion of DNA extracted from thyroid and other tissues with the isochizomers Msp I and Hpa II. It was found that one of the Hpa II sites was demethylated specifically in the thyroid.
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页码:6176 / 6180
页数:5
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