TRANSPOSON ACTIVE TRANSGENIC XIPHOPHORUS

描述(由申请人提供)：这份R21提案的重点是通过构建和验证含有能够动员SB转座子的系统的转基因鱼线来增强剑鱼遗传模型的科学能力。至少将产生两个转SB转座子的剑尾鱼品系：1)含有SB转座子的剑尾鱼JP163A及其相关报告基因的表达；2)具有相同遗传背景的第二个品系，其编码SB转座酶的基因由一个强大的组成型启动子控制。在没有转座酶蛋白的情况下，SB转座子本身不能转座。因此，这两个转基因品系的杂交有望导致转座子介导的插入突变的爆发。一旦被诱变，这些半合子转座酶/转座子动物将被用作物种间杂交的亲本。没有遗传转座酶的F1种间杂交动物(即50%的F1后代)将被计分，以调节几个复杂多基因性状的表型表达。由于剑鱼物种的多样性，将要建立的转基因系统将在遗传力方面具有无与伦比的优势，以确定涉及复杂多基因性状的候选基因，然后使用经典的遗传方法进行确认。

DESCRIPTION (provided by applicant): This R21 proposal is focused on enhancing the scientific capabilities of the Xiphophorus genetic model by construction and validation of transgenic fish lines harboring a system capable of mobilizing the SB transposon. At least two transgenic Xiphophorus lines will be produced: 1) Xiphophorus maculatus Jp 163 A containing the SB transposon with associated reporter gene expression; and 2) a second line in the same genetic background containing the gene encoding SB transposase under control of a strong constitutive promoter. The SB transposon itself cannot transpose in the absence of the transposase protein. Thus, interbreeding these two transgenic lines is expected to result in a burst of transposon mediated insertional mutagenesis. Once mutagenized, these hemizygous transposase/transposon animals will be used as parents in interspecies crosses. F1 interspecies hybrid animals that have not inherited the transposase (i.e. 50% of the F1 offspring) will be scored for modulation of phenotypic expression for several complex multigenic traits. Due to the diversity of Xiphophorus species, the transgenic system to be produced will be unparalleled in genetic power to identify candidate genes involved in complex multigenic traits that may then be confirmed using classical genetic approaches.