TRANSPOSON ACTIVE TRANSGENIC XIPHOPHORUS

描述（由申请人提供）：本R21提案的重点是通过构建和验证含有能够调动SB转座子的系统的转基因鱼系来增强剑鱼遗传模型的科学能力。至少将培育出两种转基因剑鱼系：1)含有SB转座子并表达相关报告基因的斑状剑鱼Jp 163 A；2)具有相同遗传背景的第二条序列包含编码SB转座酶的基因，该基因受强组成型启动子控制。在没有转座酶蛋白的情况下，SB转座子本身不能转座。因此，这两种转基因系的杂交将导致转座子介导的插入突变的爆发。一旦诱变，这些半合子转座酶/转座子动物将用作种间杂交的亲本。没有遗传转座酶的F1种间杂交动物（即50%的F1后代）将被评分为几种复杂的多基因性状的表型表达调节。由于剑鱼物种的多样性，要生产的转基因系统将具有无与伦比的遗传能力，可以识别涉及复杂多基因性状的候选基因，然后可以使用经典遗传方法进行确认。

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.