TRANSPOSON MUTAGENESIS OF NUCLEAR POLYHEDROSIS VIRUSES

转座是一种遗传重排的形式， 原核生物和真核生物。 它可能涉及重组 可能在许多类型的基因组重排中起作用的机制 包括T细胞受体和免疫球蛋白基因重排，基因 扩增、基因破坏和癌基因激活。 后果 转座子动员的进化高等生物和/或 真核病毒的数量可能很大。 核基因的FP突变体 多角体病毒（NPVs）提供了一个独特的实验系统， 转座子运动关键方面的分子分析。 许多这些 自发突变体是通过宿主转座子插入 病毒基因组的500 bp区域称为FP基因座。 突变 表型导致独特的斑块形态和显性 突变体在细胞培养物中繁殖期间的扩增。 这些 这些特性可用于开发体外转座测定 使用克隆的lacZ融合基因标记的转座子和靶病毒DNA。 本研究将集中在粉纹夜蛾转座子的分析上 显示靶位点特异性和插入位点的IFP 2和TFP 3 两个密切相关的NPV的FP基因座内的偏好。 的影响 在病毒靶位点或转座子末端内的突变， 将研究这些元件的插入和切除。 的能力 在分子水平上分析转座子的位点特异性插入， 这个系统的特点。 我们还将引入重组 携带不同P元件构建体的NPV在果蝇细胞中的表达 培养物和胚胎来分析转座子载体化的潜力 在不同物种之间传播 PI在以下方面做出了重大贡献： 了解FP突变体的遗传学，现在准备开发这种 一个独特的系统作为真核转座的实验模型 现象。 PI已经完成了一个为期三年的赠款，并开始 五年的持续补助金，不应该是 被认为是一个成熟的调查员。 该奖项将允许PI 直接努力建立他的实验室，作为研究的重点， 转座子遗传学 大学致力于发展一个充满活力的 分子遗传学的研究项目 私家侦探是第一个 分子遗传学家，在很大程度上负责 为研究计划的进一步发展。 这个奖项将 提高PI的能力，在培养研究生中发挥更直接的作用 学生和参与新教师的研究发展 在这个项目中。

Transposition is a form of genetic rearrangement common among both prokaryotic and eukaryotic organisms. It likely involves recombinational mechanisms that are probably active in many types of genome rearrangements including T-cell receptor and immunoglobulin gene rearrangements, gene amplification, gene disruption, and oncogene activation. The consequences of transposon mobilization for the evolution of higher organisms and/or eukaryotic viruses could be substantial. The FP mutants of nuclear polyhedrosis viruses (NPVs) provide a unique experimental system for molecular analysis of key aspects of transposon movement. Many of these spontaneous mutants arise through the insertion of host transposons within a 500 bp region of the viral genome called the FP locus. The mutant phenotype results in a distinctive plaque morphology and dominant amplification of the mutants during propagation in cell cultures. These properties can be exploited in developing an in vitro transposition assay using cloned, lacZ fusion genetagged transposons and target viral DNAs. The proposed research will focus on analysis of Trichoplusia ni transposons IFP2 and TFP3 that exhibit target site specificity and insertion site preference within the FP locus of two closely related NPVs. The effect of mutations at viral target sites or within transposon termini on the insertion and excision of these elements will be studied. The ability to analyze site-specific insertion of transposons at the molecular level is a distinguishing feature of this system. We will also introduce recombinant NPV carrying various P-element constructs into Drosophila melanogaster cell cultures and embryos to analyze the potential for vectoring of transposons between species by viruses. The PI has made significant contributions in understanding the genetics of FP mutants and is now ready to develop this unique system as an experimental model for eukaryotic transposition phenomena. The PI has completed one three year grant and is beginning a five year continuation grant on this subject, and should not yet be considered an established investigator. This award will allow the PI to direct efforts towards establishing his lab as a focus of research on transposon genetics. The University is committed to developing a vigorous research program in Molecular Genetics. The PI is the first tenured Molecular Geneticist at this institution and is responsible in large part for the further development of the research program. This award will enhance the PIs ability to take a more direct role in training graduate students and participating in the research development of the newer faculty in this program.