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New Tools for Investigating the Comparative Biology of Caenorhabditis briggsae and C. elegans

机译:研究秀丽隐杆线虫和秀丽隐杆线虫的比较生物学的新工具

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摘要

Comparative studies of Caenorhabditis briggsae and C. elegans have provided insights into gene function and developmental control in both organisms. C. elegans is a well developed model organism with a variety of molecular and genetic tools to study gene functions. In contrast, there are only very limited tools available for its closest relative, C. briggsae. To take advantage of the full potential of this comparative approach, we have developed several genetic and molecular tools to facilitate functional analysis in C. briggsae. First, we designed and implemented an SNP-based oligonucleotide microarray for rapid mapping of genetic mutants in C. briggsae. Second, we generated a mutagenized frozen library to permit the isolation of targeted deletions and used the library to recover a deletion mutant of for use as a transgenic marker. Third, we used the mutant in ballistic transformation and generated fluorescently labeled strains that allow automated lineaging and cellular resolution expression analysis. Finally, we demonstrated the potential of automated lineaging by profiling expression of , , and at cellular resolution and by detailed phenotyping of the perturbations on the Wnt signaling pathway. These additions to the experimental toolkit for C. briggsae should greatly increase its utility in comparative studies with C. elegans. With the emerging sequence of nematode species more closely related to C. briggsae, these tools may open novel avenues of experimentation in C. briggsae itself.
机译:秀丽隐杆线虫和秀丽隐杆线虫的比较研究为两种生物的基因功能和发育控制提供了见识。秀丽隐杆线虫是一种发达的模型生物,具有多种研究基因功能的分子和遗传工具。相反,只有最有限的亲戚C. briggsae可用的工具。为了充分利用这种比较方法的潜能,我们开发了几种遗传和分子工具,以促进桥花梭菌的功能分析。首先,我们设计并实现了一种基于SNP的寡核苷酸微阵列,用于快速绘制C. briggsae中的遗传突变体。其次,我们生成了诱变的冷冻文库,以允许分离目标缺失,并使用该文库回收用作基因突变标记的缺失突变体。第三,我们在突变转化中使用了该突变体,并生成了荧光标记的菌株,可以进行自动谱系和细胞分辨率表达分析。最后,我们通过分析,和在细胞分辨率下的表达以及在Wnt信号通路上的干扰的详细表型来证明自动谱系的潜力。这些对秀丽隐杆线虫的实验工具包的添加应大大提高其在秀丽隐杆线虫的比较研究中的效用。随着线虫物种的出现,与线虫的关系更加密切,这些工具可能会为线虫本身开辟新的实验途径。

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