Nucleic Acids Research, 1992, Vol. 20, No. 21 5655-5660
© 1992
MOLECULAR BIOLOGY |
The developmental regulation of the human 
-globin gene in transgenic mice employing ß-galactosidase as a reporter gene
Sir William Dunn School of Pathology, Oxford University Oxford 0X1 3RE, UK 1Department of Biochemistry, University of Sydney Sydney, NSW 2006, Australia
* To whom correspondence should be addressed
Received August 5, 1992. Accepted September 25, 1992.
We have investigated the developmental and tissue specific expression of the human embryonic 
-globin gene in transgenic mice. A construct containing 550 bp of -globin 5' flanking region, fused to a (ß-galactosidase (lacZ) reporter gene and linked to the locus control region (LCR)-like 
positive regulatory element (PRE) was employed for the production of transgenic mice. Firstly, we compared the number of live born transgenic mice containing this construct to the number of live born transgenic mice containing the entire -globin gene linked to the PRE or the ßLCR. Data showed that 12% of mice generated from eggs injected with -promoter/lacZ/PRE DNA were transgenic compared to only 2% of mice generated from eggs injected with the entire -globin gene linked to the PRE or the ßLCR. The reduced number of live born transgenic mice containing the latter constructs suggests that death of transgenic embryos, posssibly due to thalassaemia, may be occurring. X-gal staining of whole embryos containing the lacZ gene revealed that -globin promoter activity was most pronounced at 8.5–9.5 days of development and was restricted to erythroid cells. By 15 days of development, no -globin promoter activity was detected. These results suggest that the PRE can direct high level expression from the -globin promoter and that sequences required for the correct tissue and developmental specific expression of the human -globin gene are present within 550 bp's of 5' flanking region. Sequences within the body of the -globin gene or 3' of the cap site do not appear to be necessary for correct -globin developmental requlation.
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