科研之友
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摘要
Caenorhabditis briggsae as a companion species for Caenorhabditis elegans has played an increasingly important role in study of evolution of development and genome and gene regulation. Aided by the isolation of its sister spices, it has recently been established as a model for speciation study. To take full advantage of the species for comparative study, an effective transgenesis method especially those with single-copy insertion is important for functional comparison. Here, we improved a transposon-based transgenesis methodology that had been originally developed in C. elegans but worked marginally in C. briggsae. By incorporation of a heat shock step, the transgenesis efficiency in C. briggsae with a single-copy insertion is comparable to that in C. elegans. We used the method to generate 54 independent insertions mostly consisting of a mCherry tag over the C. briggsae genome. We demonstrated the use of the tags in identifying interacting loci responsible for hybrid male sterility between C. briggsae and Caenorhabditis nigoni when combined with the GFP tags we generated previously. Finally, we demonstrated that C. briggsae tolerates the C. elegans toxin, PEEL-1, but not SUP-35, making the latter a potential negative selection marker against extrachromosomal array. @@@ Caenorhabditis briggsae has been used for comparative study against Caenorhabditis elegans over decades. Here, we improved the transposon-based methodology by incorporation of a heat shock step, leading to a much higher transgenesis efficiency and generation of 54 independent insertions over the C. briggsae genome. We demonstrated that C. elegans SUP-35 can serve as a potential negative selection marker against extrachromosomal array. The improved transgenesis methodology and the transgenic strains are expected to facilitate C. briggsae as a model for comparative study.
