Genetic analysis of the ADGF multigene family by homologous recombination and gene conversion in Drosophila.
Genetic analysis of the ADGF multigene family by homologous recombination and gene conversion in Drosophila.
复制标题
通过果蝇同源重组和基因转换对 ADGF 多基因家族进行遗传分析。
作者：
T. Doležal;M. Gazi;M. Žurovec;P. Bryant
许多果蝇基因作为多基因家族的成员存在，并且在每个家族中，成员可以是功能冗余的，使得难以通过基于表型筛选的经典诱变技术来鉴定它们。我们已经解决了这个问题，在一个新的家庭的六个腺苷脱氨酶相关的生长因子（ADGF）的遗传分析。我们使用末端靶向将突变引入六个ADGF基因中的五个，利用五个家族成员由三个基因簇和两个基因簇编码的事实。我们使用两种靶向构建体将功能丧失突变引入所有五个基因，以及分离多个突变的不同组合，而不依赖于表型结果。结果表明：（1）利用末端打靶技术破坏基因簇是可能的;（2）基因转换技术，通常被认为是基因打靶中的一个复杂问题，可以用来帮助在一个单一的筛选程序中恢复不同的突变体组合;（3）通过诱导双链断裂将重复减少到单个拷贝，这可以通过以下方法得到更好的解释：链退火机制比通过重复序列之间的简单交换更重要;和（4）最丰富表达的家族成员（ADGF-A）的功能丧失导致突变幼虫中脂肪体的分解和黑色素瘤的发展。
Many Drosophila genes exist as members of multigene families and within each family the members can be functionally redundant, making it difficult to identify them by classical mutagenesis techniques based on phenotypic screening. We have addressed this problem in a genetic analysis of a novel family of six adenosine deaminase-related growth factors (ADGFs). We used ends-in targeting to introduce mutations into five of the six ADGF genes, taking advantage of the fact that five of the family members are encoded by a three-gene cluster and a two-gene cluster. We used two targeting constructs to introduce loss-of-function mutations into all five genes, as well as to isolate different combinations of multiple mutations, independent of phenotypic consequences. The results show that (1) it is possible to use ends-in targeting to disrupt gene clusters; (2) gene conversion, which is usually considered a complication in gene targeting, can be used to help recover different mutant combinations in a single screening procedure; (3) the reduction of duplication to a single copy by induction of a double-strand break is better explained by the single-strand annealing mechanism than by simple crossing over between repeats; and (4) loss of function of the most abundantly expressed family member (ADGF-A) leads to disintegration of the fat body and the development of melanotic tumors in mutant larvae.
影响因子：
10.5
作者：
Rong, YKS;Titen, SW;Golic, KG
通讯作者：
Golic, KG
果蝇 DNA 断裂的有效修复：单链退火和与其他修复途径竞争的证据。
影响因子：
3.3
作者：
Preston,ChristineR;Engels,William;Flores,Carlos
通讯作者：
Flores,Carlos
影响因子：
56.9
作者：
Rong, YS;Golic, KG
通讯作者：
Golic, KG