Conversion of RAPD markers linked to cold tolerance to scar markers in common carp
GAO Jun-sheng, SUN Xiao-wen, LIANG Li-qun
1. Heilongjiang River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Harbin 150070, China; 2. School of Life Science and Technology, Dalian Fisheries Univ. , Dalian 116023, China
Abstract: RAPD -PCR on DNA samples were performed with 300 random primers in Heilongjiang wild common carp （Cyprinus carpio）（ cold tolerance）, cold -resistance strain of red purse carp （C. carpio var. ） （cold tolermace） , red purse carp （cold sensitive）, Boshi carp （C. pellegini pellegini）（ cold sensitive）, and cross F2 of cold - resistance strain of red purse（ ♂ ） and Boshi carp（ ♀ ）. Both AL04 986 mad AR02 788 were identified to be molecular markers which were associated with cold tolerance traits of common carp. Also, specific fragments were extracted and purified from four primers above. The purified products were then inserted into pMD18 -T vector which were transformed to competent Escherichia coli DH 5α for sequencing. According to the sequences, three pairs of specific primers, SCAL04L/SCAL04R1, SCAL04L/SCAL04R2 and SCAR02L/SCAR02R, were designed, in which SCAR02L/SCAR02R was used for inversion of the difference of cold tolerance in cross F2 of cold - resistance strain of red purse and Boshi carp, and its optimal annealing temperature was 57 ℃. The PCR analysis showed that SCAR markers were associated with cold - resistance traits, which could be used for the identification of hybrid to cold tolerance traits of common carp. Unfortunately the conversed SCAR markers from AL04 986 could not reproduce the polymorphic AL04 986. The reasons for that were also discussed.
2007, Vol. 22 
