绵羊(ATAG)n四碱基微卫星位点的筛选及其在亲子鉴定中的应用

doi:10.16431/j.cnki.1671-7236.2021.07.024

Abstract

Abstract: The purpose of this study was to select the tetrameric microsatellite markers with high polymorphism,and to establish the paternity testing system of sheep.The Small-tail Han sheep was the main research object in this study,and fifty-three (ATAG)_n repeat microsatellite sites were selected according to the sheep reference genome sequence and a total of 30 microsatellite loci which had stable amplification and high polymorphism information were developed based on genotyping data in Small-tail Han sheep.The genotyping results of 30 loci showed that 253 alleles were obtained from the 30 markers,the average number of alleles was 8.433,the number of alleles was all greater than 5,the polymorphic information content (PIC) varied from 0.566 to 0.898,the calculated observed heterozygosity (Ho) ranged from 0.548 to 0.903 and the expected heterozygosity (He) ranged from 0.631 to 0.921 with a mean of 0.776.The Hardy-Weinberg equilibrium analysis revealed that all 30 sites were in genetic equilibrium.Then,22 of 30 microsatellite loci were selected according to the amplification efficiency of PCR for the calculation of parental exclusion probability,and the combinations were performed by increasing the number of bits from high to low according to the size of the PIC.The results showed that when the genotypes of the two parents were unknown,the cumulative probability of exclusion (CPE) could reach more than 99.99% when the number of marker sites was 15,and the NE-1P varied from 0.321 to 0.663.Further,16 Small-tail Han sheep with genealogical records were tested by using the established paternity test system,the results showed that 4 families with high confidence were identified,and the identification results were in complete agreement with pedigree records.This experiment laid an important foundation for the construction of sheep molecular genealogy,paternity test and the development of sheep breeding.

Key words: sheep; molecular genetic marker; tetrameric microsatellite markers; paternity analysis; genotyping

CLC Number:

S826.8⁺9

HU Mingyue, LIU Zhengxi, ZHAO Zhongli, CAO Yang, MA Huihai, YAN Shouqing. Isolation of (ATAG)_n Tetrameric Microsatellite Loci and Its Application on Paternity Analysis in Sheep[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(7): 2484-2494.

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Isolation of (ATAG)_n Tetrameric Microsatellite Loci and Its Application on Paternity Analysis in Sheep

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Isolation of (ATAG)n Tetrameric Microsatellite Loci and Its Application on Paternity Analysis in Sheep

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Isolation of (ATAG)_n Tetrameric Microsatellite Loci and Its Application on Paternity Analysis in Sheep