Michiels F, Gasch A, Kaltschmidt B, Renkawitz-Pohl R
Gentechnologische Arbeitsgruppen am MPI für Biochemie, Martinsried, FRG.
EMBO J. 1989 May;8(5):1559-65. doi: 10.1002/j.1460-2075.1989.tb03540.x.
To analyze the regulation of gene expression during male germ cell development, we investigated the testis-specific expression of the Drosophila beta 2 tubulin gene. Germ line transformation experiments with the upstream region of the D.melanogaster beta 2 tubulin gene fused to the Escherichia coli lacZ gene resulted in the correct tissue specific expression of the reporter gene. Furthermore, we showed that the upstream sequences of the beta 2 tubulin gene of the distantly related species D.hydei can drive the expression of the lacZ gene testis specifically in D.melanogaster flies. A detailed deletion analysis showed that 53 bp of upstream and 23 bp (D.melanogaster) or 29 bp (D.hydei) of leader sequences are sufficient to confer tissue specificity. The short promoter regions contain a 14 bp motif at identical positions in both species, which acts as a position-dependent promoter element. In vitro mutagenesis and subsequent germline transformation experiments revealed that this sequence is the only element necessary for the testis-specific transcription of the beta 2 tubulin gene in Drosophila.
为了分析雄性生殖细胞发育过程中的基因表达调控,我们研究了果蝇β2微管蛋白基因在睾丸中的特异性表达。将黑腹果蝇β2微管蛋白基因的上游区域与大肠杆菌lacZ基因融合进行种系转化实验,结果报告基因实现了正确的组织特异性表达。此外,我们发现远缘物种海德氏果蝇β2微管蛋白基因的上游序列能够驱动lacZ基因在黑腹果蝇睾丸中特异性表达。详细的缺失分析表明,上游53 bp以及23 bp(黑腹果蝇)或29 bp(海德氏果蝇)的前导序列足以赋予组织特异性。这两个物种的短启动子区域在相同位置含有一个14 bp的基序,它作为一个位置依赖性启动子元件发挥作用。体外诱变及随后的种系转化实验表明,该序列是果蝇β2微管蛋白基因睾丸特异性转录所必需的唯一元件。