Suppr超能文献

在果蝇黑腹果蝇的幼虫、蛹和成虫阶段中的内源性β-半乳糖苷酶活性表明在lacZ融合基因研究中需要谨慎。

Endogenous beta-galactosidase activity in the larval, pupal, and adult stages of the fruit fly, Drosophila melanogaster, indicates need for caution in lacZ fusion-gene studies.

作者信息

Schnetzer J W, Tyler M S

机构信息

Department of Zoology, University of Maine, Orono 04469-5751, USA.

出版信息

Biol Bull. 1996 Apr;190(2):173-87. doi: 10.2307/1542537.

Abstract

Beta-galactosidase activity is known to exist in Drosophila melanogaster, but a detailed analysis of the tissue-specific patterns of activity has not previously been reported. Such an analysis is of particular interest because Drosophila is commonly used for making transformants that carry fusion genes in which the E. coli beta-galactosidase gene, lacZ, is used as a reporter gene. When these transformants are analyzed for beta-galactosidase activity by using chromogen X-gal staining, the method does not distinguish true fusion-gene activity from endogenous beta-galactosidase activity or from the beta-galactosidase activity of bacterial contaminants. Therefore, detailed maps of endogenous beta-galactosidase activity in this organism would help to prevent errors in data interpretation and would indicate which stages were most appropriate for experiments with the lacZ transformants. We have constructed such maps by applying X-gal staining methods to serial frozen sections and whole mounts of larval, prepupal, pupal, and adult stages of D. melanogaster reared under axenic conditions. Results showed endogenous beta-galactosidase activity in a variety of organs including the larval intestine, spiracles, lymph glands, cellular epidermis, and eye-antenna imaginal discs; the pupal cellular epidermis, lymph glands, imaginal tissues, fat body, and spiracle; and the adult pericardial cells, thoracic nephrocytes, ventriculus, and reproductive system. The good correlation between staining and metamorphic remodeling and phagocytic activity indicates that endogenous beta-galactosidase is physiologically interesting.

摘要

已知β-半乳糖苷酶活性存在于黑腹果蝇中,但此前尚未有关于其组织特异性活性模式的详细分析报道。这样的分析特别有意义,因为果蝇常用于制备携带融合基因的转化体,其中大肠杆菌β-半乳糖苷酶基因lacZ用作报告基因。当通过使用显色剂X-gal染色分析这些转化体的β-半乳糖苷酶活性时,该方法无法区分真正的融合基因活性与内源性β-半乳糖苷酶活性或细菌污染物的β-半乳糖苷酶活性。因此,该生物体中内源性β-半乳糖苷酶活性的详细图谱将有助于防止数据解释中的错误,并指出哪些阶段最适合进行lacZ转化体的实验。我们通过将X-gal染色方法应用于在无菌条件下饲养的黑腹果蝇幼虫、蛹前期、蛹期和成虫期的连续冰冻切片和整体标本,构建了这样的图谱。结果显示,在多种器官中存在内源性β-半乳糖苷酶活性,包括幼虫肠道、气门、淋巴腺、细胞表皮和眼触角成虫盘;蛹期的细胞表皮、淋巴腺、成虫组织、脂肪体和气 门;以及成虫的心包细胞、胸肾细胞、心室和生殖系统。染色与变态重塑和吞噬活性之间的良好相关性表明内源性β-半乳糖苷酶在生理上具有重要意义。

文献AI研究员

20分钟写一篇综述,助力文献阅读效率提升50倍。

立即体验

用中文搜PubMed

大模型驱动的PubMed中文搜索引擎

马上搜索

文档翻译

学术文献翻译模型,支持多种主流文档格式。

立即体验