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Open Access Article

Scientific Development Research . 2022; 2: (1) ; 46-52 ; DOI: 10.12208/j.sdr.20220012.

Construction and Identification of Adck Transgenic Flies
Adck转基因果蝇品系的构建与鉴定

作者: 龙芸*,

广州医科大学 广东广州

*通讯作者: 龙芸,单位:广州医科大学 广东广州;

发布时间: 2022-06-14 总浏览量: 26

摘要

目的 Adck转基因果蝇品系的构建与鉴定。方法 蛋白质磷酸化是指由蛋白质激酶催化的把ATP的磷酸基转移到底物蛋白质氨基酸残基(丝氨酸、苏氨酸、酪氨酸)上的过程,或者在信号作用下结合GTP,是生物体内一种普通的调节方式,在细胞信号转导的过程中起重要作用。蛋白质磷酸化是调节和控制蛋白质活力和功能的最基本、最普遍,也是最重要的机制。Adck是果蝇的蛋白编码基因,其蛋白含有aarF结构域,目前Adck在果蝇中的功能还不清楚。为进一步研究Adck基因的功能,我们利用PhiC31整合酶介导的定点转基因系统构建带HA标签的转基因果蝇。结果 成功构建Adck转基因果蝇。结论 本实验以果蝇的cDNA 为模板扩增Adck CDS区,同时双酶切载体pUAST-attB,进行重组连接,并对阳性克隆进行PCR鉴定及测序;将测序正确的质粒注射到果蝇的胚胎细胞,经过一系列的杂交,筛选出转基因果蝇,经其自交可获得纯合的Adck转基因果蝇。

关键词: Adck基因;转基因果蝇

Abstract

Objective Construction and Identification of Adck Transgenic Flies.
Methods Protein phosphorylation refers to the process of transferring the phosphoryl group of ATP to the amino acid residues (serine, threonine, tyrosine) of the substrate protein catalyzed by protein kinase, or binding GTP under the action of signal, which is a common regulating mode in organisms and plays an important role in the process of cell signal transduction. Protein phosphorylation is the most basic, common and important mechanism for regulating and controlling protein activity and function.Adck is a protein-coding gene of Drosophila. The protein encoded by Adck contains aarF domain. At present, the function of Adck in Drosophila is not clear. To further study the function of Adck, we use PhiC31 integrase-mediated site-specific transgenic system to construct transgenic flies with HA tag.
Results Successful construction of Adck transgenic flies.
Conclusion   This experiment use cDNA as the template to amplify Adck CDS, and digest vector pUAST-attB with two enzymes, and then ligase these two fragments. After that we identified positive clones by PCR. Then we injected the right plasmid into the germ cells of Drosophila melanogaster. After a series of crosses,we finally get Adck transgenic flies.

Key words: Adck; Transgenic flies

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引用本文

龙芸, Adck转基因果蝇品系的构建与鉴定[J]. 科学发展研究, 2022; 2: (1) : 46-52.