Microsatellite Cloning And Screening Of The AFLP Markers Linked To The Sex Locus Of Acipenser Schrenckii

Amur sturgeon (Acipenser schrenckii) is a cartilaginous fish (Chondrichthyes) Acipenseriformes (Acipenseriformes) sturgeon division (Aeipenseridac) is a sturgeon (Acipenser Linnaeus) of fish, unique to the Heilongjiang River sturgeon, with individual, long life, survival rate High, less disease, grow faster and so is the most economic value in beluga sturgeon of rare fish, artificial breeding prospect. This study analyzed by AFLP technology belong to Siberia sturgeon farming groups of genetic structure, 7 primer pairs were used to amplify 24 Acipenser baeri genomic DNA samples by AFLP method.There are total of 234 loci detected with 24³9 loci per primer pair, in which 116 loci are polymorphic with polymorphisms of 48.83%.The mean Nei's genetic diversity index(H) is found 0.4297, the mean Shannon's information index(I) 0.1233, showing a relative lower genetic polymorphism. The mean genetic distance among individuals is found 0.1872.Microsatellite enrichment by magnetic beads is a fast and efficient method for isolating these molecular genetic markers. A part genomic library was created and hybridized with a [γ32^P] labeled probe (GA)₁₂,(CAT)₈,(AGAT)₆. Among the 3320 clones, 1200 sequencing sent successfully sequenced 1136, by 1048(rate 92.25%) clones containing microsatellite sequences. There were 1529 microsatellite DNA in all and repeated 10 times or more the number of microsatellites accounted for 65.24%.85.74% microsatellites were perfect repeats, 3.60% imperfect repeats and 10.66% compound perfect repeats. This allowed us to designed 299 pairs of primers with the software Primer 3.0. 200 pairs of primers were composted and screened. 116 pairs amplified special PCR products in the population of Amur sturgeon and 16 pairs were polymorphism in the population of Amur sturgeon. Also selected two female specific molecular markers HLJSX178 and HLJSX209.The microsatellite markers isolated here will become very useful genetic tools in the population diversities analyze, conservation biology and strains identification.Genomic DNA was extracted from fins of female and male Amur sturgeon using high concentrated salty-PCI(Pheno1,Chloroform,Isoamylalcoho1). The amplified fragment-length polymorphism (AFLP) technique was used to identify sex-specific markers in Amur sturgeon.Sixty four pairs of primers of the 8 EcoR I-NNN and 8 Mse I-NNN combination were used to amplify the mixed male and female DNA pools, 3264 flagments were amplified and 379 candidate differential DNA fragments were obtatned. This study was the first attempt to find sex-specific markers in Acipenser schrenckii.Despite the failure to find such markers, these data offer useful information for further studies targeting similar goals.