Microsatellite Markers Development And Polymorphism Analysis In The Laboratory Red Carp Carp C1HD

Objective: To screen out the microsatellite markers and to analysis microsatellite polymorphisms in the laboratory red carp crucian C1 HD.Methods: First,Micro Satellite identificantion tool was used to predict the SSR information of the whole genome of red carp crucian.Based on the genome-wide information of red carp crucian,110 pairs of microsatellite primers were designed using Primer Premier 5.0 software,and the optimal annealing temperature of SSR-PCR for each microsatellite primer was determined by gradient PCR.According to the sequence of microsatellite locus in the genome of red common carp,the genomic DNA of 10 laboratory red carp crucian C1 HD and 10 comon red carp crucian were amplified by PCR,8% non-denaturing polyacrylamide gel electrophoresis was used to screening out microsatellite DNA markers from laboratory red carp crucian C1 HD.Sequencing analysis was performed on the specific bands of the laboratory red carp crucian C1 HD.40 laboratory red carp crucian C1 HD were analyzed by 5 Pairs of laboratory red carp crucian C1HD-specific Microsatellite Primers.The genetic quality of the laboratory red carp crucian C1 HD,Gene frequency(P),Expected heterozygosity(UHe),Expected homozygosity(Hom),and Polymorphism information content(PIC)were tested with the screened SSRs molecular markersbased on <Laboratory Fish Genetic Quality Control of Laboratory Red Carp C1 HD Strain >.Results: 110 pairs of SSR-PCR microsatellite primers were amplified.According to the specificity of its band,24 pairs of microsatellite primers were screened out.Using these 24 pairs of microsatellite primers for SSRPCR,the laboratory red carp crucian C1 HD can be distinguished from the comon red carp crucian,so it can be used as an symbolic microsatellite primer for thlaboratory red carp crucian C1 HD of common red carp crucian.A total of 11 sites were amplified by PCR amplification of 40 laboratory red carp crucian C1 HD with 5 pairs of laboratory red carp crucian C1 HD specific microsatellite primers.The average unbiased expected heterozygosity(UHe),average expected homozygosity(Hom),and average polymorphism information content(PIC)for laboratory red carp crucian C1 HD were 0.081,0.919,and 0.076,respectively.The results of the sequencing of the specific bands of the microsatellite primer22 indicate that the sequence has a high similarity to the CAPN3 gene sequence.Conclusions: 24 pairs of microsatellite-specific primers can be used as microsatellite markers for laboratory red carp crucian C1 HD,and the laboratory red carp crucian C1 HD can be distinguished from common red carp crucian.The average expected homozygosity(Hom)of the 40 individuals of the second generation of the laboratory red carp crucian C1 HD was 0.919,indicating a higher degree of gene homozygosity and could be used as an inbred strain.