Salivary Gland Genetic Markers

唾液腺遗传标记

• 批准号: 8917985
• 负责人: Steven Beckendorf
• 金额: $ 18万
• 依托单位: University of California-Berkeley
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-07-15 至 1992-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=8917985&HistoricalAwards=false
• 关键词: Salivary; Gland; Genetic; Markers

The embryonic development of the Drosophila salivary glands provides several advantages for analysis of the genes involved in determination and differentiation. In the ectoderm of the germ band extended embryo, the glands begin development at a defined position which suggests the involvement of several known pattern regulating genes. Differentiation then occurs within a few hours, implying that there are few steps leading from commitment to differentiation. This proposal suggests methods to identify genes acting in the salivary glands at this early stage and to order them into a pathway for salivary gland development. Salivary gland marker genes will be identified using P-lacZ transposons that express beta-galactosidase in tissue specific patterns that include the salivary glands. Embryos homozygous for lethal transposon inserts will be examined for phenotypes that alter salivary gland development. For non-lethal inserts, mutations will be induced by imprecise transposon excision to test whether an adjacent gene is required for the development of salivary glands. For some inserts that are shown to be within or near to a gene that is important for salivary gland development, regions flanking the insert will be cloned by plasmid rescue and these regions searched for embryonic salivary gland transcripts. To get clues to gene function, exonic regions will be sequenced and examined for homology with known genes. Finally, mutations in required genes will be examined for effects on beta-galactosidase expression in several enhancer trap lines. Results form these tests will help define a pathway for embryonic salivary gland development. %%% Investigations of the genes that form patterns in early drosophila development have been spectacularly successful in solving the longstanding riddle of how a single fertilized egg cell develops the intricate and regular pattern that characterizes the mature embryo. This proposal is an attempt to extend those studies downstream to the actual formation of a differentiated organ, the salivary gland.

果蝇唾液腺的胚胎发育为分析参与测定和分化的基因提供了多种优势。在种带延伸胚胎的外胚层中，腺体在一个确定的位置开始发育，这表明几个已知的模式调节基因的参与。然后，差异化会在几个小时内发生，这意味着从承诺到差异化只需几个步骤。该提案提出了一些方法来识别早期阶段在唾液腺中起作用的基因，并将它们排列成唾液腺发育的途径。 将使用 P-lacZ 转座子来鉴定唾液腺标记基因，该转座子以包括唾液腺在内的组织特异性模式表达 β-半乳糖苷酶。将检查致命转座子插入的纯合胚胎是否改变唾液腺发育的表型。对于非致死插入，将通过不精确的转座子切除诱导突变，以测试唾液腺的发育是否需要相邻基因。对于某些被证明位于对唾液腺发育重要的基因内或附近的插入物，插入物侧翼的区域将通过质粒救援来克隆，并且这些区域搜索胚胎唾液腺转录物。 为了获得基因功能的线索，将对外显子区域进行测序并检查其与已知基因的同源性。最后，将检查所需基因的突变对几个增强子捕获系中β-半乳糖苷酶表达的影响。这些测试的结果将有助于确定胚胎唾液腺发育的途径。 %%% 对果蝇早期发育中形成模式的基因的研究取得了巨大成功，解决了单个受精卵细胞如何发育出成熟胚胎特征的复杂而规则的模式这一长期存在的谜团。该提议试图将这些研究扩展到下游，直至分化器官——唾液腺——的实际形成。