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HEREDITAS ›› 2010, Vol. 32 ›› Issue (1): 67-72.doi: 10.3724/SP.J.1005.2010.00067

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Construction of genome BAC library for single Branchiostoma belcheri individual

ZHANG Ju-Yong1,HUANG Sheng-Feng2,WANG Wei3,XU An-Long2,WANG Yi-Quan1   

  1. 1. School of Life Sciences, Xiamen University, Xiamen 361005, China; 2. College of Life Sciences, Sun Yat-Sen (Zhongshan) University, Guangzhou 510275, China; 3. The Third Institute of Oceanography, State Oceanic Administration, Xiamen 361003, China
  • Received:2009-08-27 Revised:2009-10-10 Online:2010-01-20 Published:2010-01-15
  • Contact: Wang Yiquan E-mail:wangyq@xmu.edu.cn

Abstract:

As one of the closest living invertebrate relatives of vertebrates, amphioxus (subphylum Cephalochordata) occupies a key position in animal evolution and is becoming the best available proxy and model animal for studying the last common ancestor of all chordates, especially vertebrates. As long-term continuous culturing of amphioxus in laboratory became reliable, for pushing this animal to be a more successful model system, whole-genome sequencing of one or more species derived from this branch will be another urgent issue needed to address. In the present study, we described the construction and characterization of a bacterial artificial chromosome (BAC) library, using a single individual of Chinese amphioxus (Branchiostoma belcheri). High quality genomic DNA extracted from the spermary was partially digested with EcoRⅠand EcoRⅠmethylase. Desirable DNA fragments were isolated by pulsed field gel electrophoresis (PFGE), ligated to linearized and phosphorylased carrier pCC1BAC, and then transformed to Escherichia coli EPI300. The constructed library consists of 44 706 clones with the average insert fragment size around 80 kb as estimated by PFGE. The representation of the library is about 9 equivalents to the amhioxus genome. These results indicate that the BAC library will be useful for functional genomic studies and facilitate the whole-genome sequencing of Chinese amphioxus.

Key words: amphioxus, Cephalochordata, Branchiostoma belcheri, genomic library, BAC