Abstract: Microsatellite markers were used to make marker regression analysis on activity of lactate dehydrogenase based on double pseudo-testcross strategy using Windows Map Manager2.0 software. The parents that came from the cross between progenies of Hebao-cold tolerance red carp and Barbless carp and F₂ progenies were used as segregating popula-tions. For maker regression, a total of 12 markers associated with activity of lactate dehydrogenase were significant at P<0.05 and HLJE222 was significant at P<0.01. The variance explained by these loci, ranged from 4.00% to 10.00%. Lo-cus HLJE222 was closely linked to the gene related to activity of lactate dehydrogenase of common carp. For further identi-fication, EST-SSR markers were used to screen the protein and nucleotide database using bioinformatics tools in order to find the homologies. High sequence similarities of HLJE222 marker were observed with the nucleotide sequence of DAZ associated protein 1mRNA of zebrafish(94%), and protein sequence of DAZ associated protein 1(97%). DAZ protein is one of the short chain dehydrogenases, which is an important enzyme in the process of glucose metabolism in the organisms. This family contains a wide variety of dehydrogenases. This indicates that locus HLJE222 was closely linked to the gene associated with activity of lactate dehydrogenase of common carp.