Xenbase Image ID: 130423
Figure 1. Somatic remobilization of a single copy Tol2 transposon in Xenopus tropicalis. (a) Schematic of the micro-injection remobilization strategy. Embryos were injected with synthetic Tol2 transposase mRNA at the one-cell stage and allowed to develop to swimming tadpole stage before genomic DNA was harvested for analysis of the integration sites by EPTS LM-PCR. (b) Sequence of the Tol2 integration sites identified in the 'remobilized' tadpoles. Two independent single transposon integration sites (10M and 12M) were used to demonstrate somatic remobilization of Tol2 transposons in Xenopus tropicalis. The parental integration sites are shown in bold. Examples of novel re-integration sites, indicating remobilization of the parental transposon, are listed below the parental site and are labelled to identify the tadpole and clone number (for example, 10M-tadpole number-clone number). Multiple remobilization events could be identified in individual tadpoles, tadpole 10M-1 had four novel integration events, suggesting that multiple independent remobilizations had occurred in discrete blastomeres during early development. All tadpoles analyzed (18/18 = 100%) had at least one novel re-integration event. The parental transposon could be remobilized from either male or female Tol2XIG F1 transgenic donor animals. The genomic sequence flanking the 5'-end of the transposon is shown in capital letters and the eight base pair target site duplication (TSD) is underlined. The transposon sequence in depicted by lower case italics text. The transposon flanking sequences were compared to the Xenopus tropicalis genome sequence (JGI assembly v4.1) to identify the scaffold identity of the integration site and to identify flanking genes; the gene nearest to the transposition site is listed in the table.
Image published in: Yergeau DA et al. (2010)
Image downloaded from an Open Access article in PubMed Central. Copyright ©2010 Yergeau et al; licensee BioMed Central Ltd.
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