Glaser R L, Wolfner M F, Lis J T
EMBO J. 1986 Apr;5(4):747-54. doi: 10.1002/j.1460-2075.1986.tb04277.x.
The tissue-specific patterns of developmental expression of hsp26-lacZ fusion genes inserted into Drosophila melanogaster by germline transformation were analyzed in several transformant lines utilizing a histochemical assay for beta-galactosidase activity on whole animals. We compared this pattern to the tissue-specific distribution of endogenous hsp26 RNA determined using hybridization of probes to RNA in situ in tissue sections. Both assays reveal that hsp26 is expressed in numerous tissues during development including spermatocytes, nurse cells, epithelium, imaginal discs, proventriculus and neurocytes. The ease and resolution of the whole-animal beta-galactosidase assay makes it particularly attractive for the elucidation of sequences involved in such complex regulation. The original hsp26-lacZ fusion gene contained 2 kb of sequence upstream of the transcription start. A construct containing only 278 bp upstream was still expressed in spermatocytes but no longer in nurse cells. In a few instances, the fusion genes were expressed in tissues for which there was no evidence for expression of the endogenous hsp26 gene. These novel patterns appear to be a result of chromosomal position since they were observed in only one or a subset of transformant lines containing identical inserts.
利用对整个动物进行β-半乳糖苷酶活性的组织化学分析,在几个转基因品系中分析了通过种系转化插入黑腹果蝇的hsp26-lacZ融合基因的组织特异性发育表达模式。我们将这种模式与使用探针与组织切片中的RNA进行原位杂交确定的内源性hsp26 RNA的组织特异性分布进行了比较。两种分析都表明,hsp26在发育过程中在许多组织中表达,包括精母细胞、滋养细胞、上皮、成虫盘、前胃和神经细胞。全动物β-半乳糖苷酶分析的简便性和分辨率使其对于阐明参与这种复杂调控的序列特别有吸引力。原始的hsp26-lacZ融合基因在转录起始上游包含2 kb的序列。一个仅包含上游278 bp的构建体仍在精母细胞中表达,但不再在滋养细胞中表达。在少数情况下,融合基因在没有证据表明内源性hsp26基因表达的组织中表达。这些新的模式似乎是染色体位置的结果,因为它们仅在包含相同插入片段的一个或一部分转基因品系中观察到。