Nucleic Acids Research, 1991, Vol. 19, No. 16 4515-4521
© 1991
MOLECULAR BIOLOGY |
Further sequence requirements for male germ cell-specific expression under the control of the 14 bp promoter element (ß2UE1) of the Drosophila ß2 tubulin gene
Genzentrum am MPI für Biochemie Am Klopferspitz 18A, D-8033 Martinsried, FRG
*To whom correspondence should be addressed at: tnstitut für Molekulargenetik, Fachbereich Biologie, Philipps-Universität-Marburg, Postfach 1929, 3550 Marburg, FRG
Received April 4, 1991. Revised July 26, 1991. Accepted July 26, 1991.
We have investigated a 14 bp promoter element (ß2UE1) that is required for testis-specific expression of the Drosophila ß2 tubulin gene. To further elucidate the role of the 14 bp element, we fused different promoter constructs to the E. coli lacZ gene and established transgenic strains with the aid of the Drosophila P-element transformation system. Germ line transformation experiments with constructs in which the element in the ß2 tubulin gene promoter was exchanged for a related sequence from the promoter region of the Drosophila ß3 tubulin gene led to a dramatic reduction in the expression of the lacZ gene in the testis. Exchanging the 14 bp promoter element for a similar sequence from the distal promoter of the Drosophila alcohol dehydrogenase gene abolished expression. This might indicate that the sequence differences between the ß2UE1 and the ß2UE1-related elements reflect functional differences between these elements. Constructs in which the ß2UE1 was fused to the hsp70 promoter revealed that testis-specific expression of a marker gene is obtained only when the element is located at the correct distance from the transcription initiation site. However, constructs in which the ß2UE1 was inserted at about the correct position (between –41 and –54 bp) upstream of a truncated ß3 tubulin gene promoter did not show any expression. By making ß2–ß3 gene promoter fusions it was found that both the region surrounding the ß3 transcription initiation site as well as the first 116 b of ß3 leader sequences independently reduce testis-specific expression. These findings suggest that the testis-specific expression of the Drosophila ß2 tubulin gene underlies a unique regulatory mechanism.
+ Present address: Department of Anatomy and Embryologie, University of Amsterdam, Meibergdreef 15, 1101 AZ Amsterdam, The Nethedands.
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