{{'Search' | translate}}
 

Tris(hydroxymethyl)aminomethane (TRIS, Trometamol) Gen-Apex® for molecular biology

三(羟甲基)氨基甲烷(TRIS,氨丁三醇)Gen-Apex for molecular biology

Company: VWR
Catalog#: 33621.26
Bio-protocol()
Company-protocol()
Other protocol()

Protein Extraction from Drosophila Embryos and Ovaries
Author:
Date:
2015-05-05
[Abstract]  Here we provide the description of protocols to efficiently obtain protein extracts from embryos and ovaries of Drosophila melanogaster. These protocols are routinely applied in our laboratory and are based on two techniques: either embryos or ovaries are homogenized using a pestle and then the soluble proteins separated by centrifugation, or embryos are individually lysed by needle manipulation. The latter technique allows the use of small embryo numbers and the selection of specific developmental stages (Guilgur et al., 2014). [摘要]  FLP / FRT系统是基于将重组酶(翻转酶-FLP)靶向指定为翻转酶识别靶标(FRT)位点的特定DNA区域的定点重组技术。最初在酿酒酵母中鉴定,酵母FLP酶及其FRT重组靶点成功转移到果蝇中的每个主要染色体臂(Golic和Lindquist,1989)。这提供了以受控方式在发育过程中在体内介导有丝分裂重组的能力[在Theodosiou和Xu(1998)中修订]。有丝分裂重组事件的受控诱导通常通过在热休克(hs)启动子的控制下表达FLP来进行。这允许在特定发育时间窗口表达高FLP水平。携带这些遗传标记的FLP / FRT染色体的菌株大大增强了我们在种系和体细胞果蝇组织中研究基因功能的能力。在这里我们描述两种不同的方案:一种用于诱导和鉴定卵巢中的纯合突变体克隆,另一种用于产生雌性种系突变体,用于分析母体对胚胎发生的影响。

Comments