Characterization of class II A and B genes in a gynogenetic carp clone

Abstract

A prerequisite for carrying out functional studies on major histocompatibility complex (Mhc) molecules of fish is the availability of genetically well-defined homozygous strains. Previously we have applied gynogenetic reproduction to generate isogenic carp, denoted clone A410. This clone has recently been demonstrated to express a single class I gene, Cyca-UA1(*)01, and in the present study two class II B and two class II A transcripts were obtained. The two class II B transcripts, Cyca-D(CB3)B and Cyca-D(CB4)B, as well as the class II A transcripts, Cyca-D(10A)A and Cyca-D(15A)A, appear to be bona fide class II transcripts, based on the presence of conserved protein characteristics of the inferred class II molecules. With the isolation of class II A sequences, representatives of all major classes of Mhc genes have been identified in the carp. To assess the relationship between the different class II genes, segregation studies, comparison of cDNA and intron 1 sequence data, and phylogenetic analyses were undertaken. These showed that the class II B transcripts, Cyca-D(CB3)B and Cyca-D(CB4)B, are derived from related, closely linked loci. In addition, these studies indicated that the previously described Cyca-DAB*01 and Cyca-DAB*02 are also closely linked, but that this linked pair segregates independently from the Cyca-D(CB3)B and Cyca-D(CB4)B loci. The class II A transcripts are most likely derived from separate loci and do not represent alleles, as they were found not to segregate in the individuals of the clone which was generated by meiogenetic gynogenesis.