Construction of a BAC library for Chinese amphioxus Branchiostoma belcheri and identification of clones containing Amphi-Pax genes.
Amphioxus is a crucial organism for the study of vertebrate evolution. Although a genomic BAC library of Branchiostoma floridae has been constructed, we report here another BAC library construction of its distant relative species Branchiostoma belcheri. The amphioxus BAC library established in present study consists of 45,312 clones arrayed in one hundred and eighteen 384-well plates. The average insert fragment size was 120 kb estimated by Pulsed Field Gel Electrophoresis (PFGE) analysis of 318 randomly selected clones. The representation of the library is about 12 equivalent to the genome, allowing a 99.9995% probability of recovering any specific sequence of interest. We further screened the library with 4 single copied Amphi-Pax genes and identified total of 26 positive clones with average of 6.5 clones for each gene. The result indicates this library is well suited for many applications and should also serve as a useful complemental resource for the scientific community.
Wang
Wei
W
Key Laboratory of the Ministry of Education for Cell Biology and Tumor Cell Engineering, School of Life Sciences, Xiamen University, China.
Xu
Huai-Liang
HL
Lin
Lu-Ping
LP
Su
Bing
B
Wang
Yi-Quan
YQ
D000818:Animals; D002681:China; D002816:Chordata, Nonvertebrate / Q000235:genetics*; D022202:Chromosomes, Artificial, Bacterial / Q000235:genetics*; D017931:DNA Primers; D016521:Electrophoresis, Gel, Pulsed-Field; D015723:Gene Library; D051761:Paired Box Transcription Factors / Q000235:genetics
D017931:DNA Primers; D051761:Paired Box Transcription Factors