The cloning of Dmc1 cDNAs and a comparative study of its expression in different ploidy cyprinid fishes

摘要

Dmrf (disrupted meiotic cDNA) is a functionally specific gene,which was firstly discovered in yeast and then found to encode a protein required for homologous chromosome synapsis during the process of meiosis.In this investigation,we cloned the partial cDNAs of Dmrf of diploid red crucian carp,Japanese crucian carp,common carp,triploid crucian carp and allotetraploid hybrids by using a pair of degenerate primers based on the conservative sequence of amino acids of the DMC1 protein in yeast,mouse and human.The full length cDNAs were then obtained by rapid amplification of cDNA ends (RACE).Our data showed that the full length cDNAs of Dmrf in the three diploid fishes are all 1375 bp long,while it is 1383 bp long in triploids and 1379 bp long in allotetraploids.And despite of the variation in length,all the cDNAs encode a protein of 342 amino acids.A high homology of 97.3% of the DMC1 protein can be drawn by comparing the amino acid sequences in the three diploids,which is also of 86%,86% and 95% similarity to human,mouse and zebrafish,respectively.A comparative study of the expression pattern of Dmrf was carried out by RT-PCR using specific primers against the same sequences of coding regions in different ploidy cyprinid fishes,from which it was showed that Dmrf was expressed only in gonads of these five kinds of fishes.The expression pattern of Dmrf in both ovaries and testes from different ploidy fishes within breeding season was also studied by Real-time PCR,and the results showed that the expression of this gene was greatly different among the three different ploidy fishes,which was the highest of triploid and lowest of allotetraploids.The histological sections data showed matured gonads of both diploid red crucian carp and allotetraploids in breeding season,although the latter demonstrated a higher maturation,and no gonadal maturation could be observed in triploids.In conclusion,we suggest that Dmrf is specifically expressed in the period of meiosis in all the ploidy cyprinid fishes and directly related with the development of gonad in a manner of ploidy-independent way.And further,the high expression of Dmrf in female triploids might be associated with abnormal meiosis and sterility.