Real time quantitative PCR system

繁殖性状在物种间经历快速变化，这种快速变化的模式也可以在对与繁殖有关的基因的DNA序列分析中发现。快速进化可以通过交配成功的性内和性间竞争以及影响受精的交配后事件来塑造。我的目标是确定导致精子发育差异、受精前精子竞争相互作用（即精子竞争）和影响受精的精子×雌性相互作用引起的适应性差异的基因。众所周知，受精成功率的差异存在遗传基础，但通常使用的方法无法识别单个基因。现在可以调查整个基因组中与适应性差异相关的基因表达差异。我的工作将有助于识别与精子发育和受精成功差异相关的基因表达模式。需要基因表达的真实的实时定量测定以通过基因表达差异的适当定量来鉴定这些基因。确定负责精子发育、精子竞争能力和受精成功差异的特定基因将有助于阐明导致不育的生殖特征、物种间生殖隔离和雄性×雌性生殖相互作用的遗传基础。在这个项目中确定的基因将用于序列相似性的比较基因组搜索，提示保守的功能和适应性变化与进化过渡。在果蝇中鉴定的基因可以作为在具有商业价值的昆虫中进行检测的候选基因，这些基因可以在实验室条件下进行测试。同样，控制果蝇精子发育的基因在人类中也有同源物（例如boule），因此果蝇精子发育转录组的表征将有助于寻找影响人类男性生育力的候选基因。

Reproductive traits have been shown to experience rapid changes among species, and such pattern of rapid change can also be found in DNA sequence analysis of genes with a role in reproduction. Rapid evolution can be shaped by intra and intersexual competition for mating success as well as postmating events that influence fertilization. My goal is to identify genes that contribute to differences in fitness caused by differences in sperm development, sperm competitive interactions prior to fertilization (i.e. sperm competition) and sperm × female interactions affecting fertilization. It is known that there is a genetic basis to differences in fertilization success, but the approach generally used provides no identification of single genes. It is now possible to survey entire genomes for differences in gene expression that associate with differences in fitness. My work shall contribute to identify genes with patterns of expression that associate with differences in sperm development and fertilization success. Real time quantitative assays of gene expression are needed to identify such genes through proper quantification of differences in gene expression. Identifying specific genes that are responsible for differences in sperm development, sperm competitive ability and fertilization success will shed light on the genetic basis of reproductive characters contributing to infertility, reproductive isolation between species and male × female reproductive interactions. Genes identified in this project will be used in comparative genomic searches of sequence similarities suggestive of conserved function and adaptive changes linked to evolutionary transitions. Genes identified in Drosophila can serve as candidates for assays in insects of commercial value that could be tested in laboratory conditions. As well, genes controlling sperm development in Drosophila have homologs in humans (e.g. boule) and so the characterization of the sperm development transcriptome in fruit flies will help in search of candidate genes affecting human male fertility.