Fosmid Library Construction and Initial Analysis o

Authors: Xubo Wang Quanqi Zhang Xiaohua Sun Yanjie Chen Teng Zhai Wei Zhuang Jie Qi Zhigang Wang

Publish Date: 2008/09/02

Volume: 11, Issue: 2, Pages: 236-242

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Abstract

Halfsmooth tongue sole Cynoglossus semilaevis Pleuronectiformes is a commercially important cultured marine flatfish in China and forms an important fishery resource but the research of its genome is underdeveloped In this study we constructed a female C semilaevis fosmid library and analyzed the fosmid end sequences to provide a preliminary assessment of the genome The library consists of 49920 clones with an average insert size of about 39 kb amounting to 323 genome equivalents Fosmid stability assays indicate that female C semilaevis DNA was stable during propagation in the fosmid system Library screening with eight microsatellite markers yielded between two and five positive clones and none of those tested was absent from the library Endsequencing of both 5′ and 3′ ends of 1152 individual clones generated 2247 sequences after trimming with an average sequence length of 855 bp BLASTN searches of the nr and EST databases of GenBank and BLASTX searches of the nr database resulted in 259 1153 and 287 1277 significant hits E  e −5 respectively Repetitive sequences analysis resulted in 523 of base pairs masked using both the Fugu and Danio databases repetitive elements were composed of retroelements DNA transposons satellites simple repeats and lowcomplexity sequences The fosmid library in conjunction with the fosmid end sequences will serve as a useful resource for largescale genome sequencing physical mapping and positional cloning and provide a better understanding of female C semilaevis genome

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