Construction of two BAC libraries from half-smooth tongue sole Cynoglossus semilaevis and identification of clones containing candidate sex-determination genes.
Half-smooth tongue sole (Cynoglossus semilaevis) is an increasingly important aquaculture species in China. It is also a tractable model to study sex chromosome evolution and to further elucidate the mechanism of sex determination in teleosts. Two bacterial artificial chromosome (BAC) libraries for C. semilaevis, with large, high-quality inserts and deep coverage, were constructed in the BamHI and HindIII sites of the vector pECBAC1. The two libraries contain a total of 55,296 BAC clones arrayed in 144 384-well microtiter plates and correspond to 13.36 haploid genome equivalents. The combined libraries have a greater than 99% probability of containing any single-copy sequence. Screening high-density arrays of the libraries with probes for female-specific markers and sex-related genes generated between 4-46 primary positive clones per probe. Thus, the two BAC libraries of C. semilaevis provided a readily useable platform for genomics research, illustrated by the isolation of sex determination gene(s).
Shao
Chang-Wei
CW
Key Laboratory for Sustainable Utilization of Marine Fisheries Resources, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, 106 Nanjing Road, Qingdao, Shandong, 266071, China.
Chen
Song-Lin
SL
Scheuring
Chantel F
CF
Xu
Jian-Yong
JY
Sha
Zhen-Xia
ZX
Dong
Xiao-Li
XL
Zhang
Hong-Bin
HB
D000818:Animals; D022202:Chromosomes, Artificial, Bacterial / Q000235:genetics*; D003001:Cloning, Molecular; D005260:Female; D005398:Fisheries / Q000379:methods*; D005412:Flatfishes / Q000145:classification / Q000235:genetics*; D005786:Gene Expression Regulation / Q000235:genetics*; D015723:Gene Library; D008297:Male; D019849:Sex Determination Processes / Q000235:genetics*