Transposon Tc1-derived, sequence-tagged sites in Caenorhabditis elegans as markers for gene mapping

被引:67
作者
Korswagen, HC
Durbin, RM
Smits, MT
Plasterk, RHA
机构
[1] NETHERLANDS CANC INST, DIV MOL BIOL, NL-1066 CX AMSTERDAM, NETHERLANDS
[2] SANGER CTR, HINXTON CB10 1RQ, CAMBRIDGE, ENGLAND
基金
英国惠康基金;
关键词
genome project; forward genetics; reverse genetics;
D O I
10.1073/pnas.93.25.14680
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
We present an approach to map large numbers of Tc1 transposon insertions in the genome of Cacnorhabditis elegans. Strains have been described that contain up to 500 polymorphic Tc1 insertions, From these we hare cloned and shotgun sequenced over 2000 Tc1 flanks, resulting in an estimated set of 400 or more distinct Tc1 insertion alleles. Alignment of these sequences revealed a weak Tc1 insertion site consensus sequence that was symmetric around the invariant TA target site and reads CAYATATRTG. The Tc1 flanking sequences were compared with 40 Mbp of a C. elegans genome sequence, We found 151 insertions within the sequenced area, a density of approximate to 1 Tc1 insertion in every 265 kb. As the rest of the C. elegans genome sequence is obtained remaining Tc1 alleles will fall into place, These mapped Tc1 insertions can serve two functions: (i) insertions in or near genes can be used to isolate deletion derivatives that have that gene mutated: and (ii) they represent a dense collection of polymorphic sequence-tagged sites, We demonstrate a strategy to use these Tc1 sequence-tagged sites in fine-mapping mutations.
引用
收藏
页码:14680 / 14685
页数:6
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