Solem Stein Tore, Jørgensen T Trond Ø
Department of Marine Biotechnology, Norwegian College of Fishery Science, Breivika 9037 Tromsø, Norway.
Dev Comp Immunol. 2002 Sep;26(7):635-47. doi: 10.1016/s0145-305x(02)00012-5.
By screening a cDNA library and analysis of DNA produced by a combined 3'RACE/5'-anchored PCR, we have isolated three isotypes of IgL in the Atlantic salmon. Two of the isotypes were homologous to rainbow trout IgL1 and L2 sequences, while the third represents a previously uncharacterised salmonid IgL. The novel type 3 CL region is homologous to spotted wolffish c1 and yellowtail sequences, while the VL region is more similar to channel catfish F class than to any other fish VL sequences. Southern analysis indicates that the gene segments of all three isotypes are organised in multiple clusters. In addition, the VL gene segments of type 3 are arranged in opposite orientation relative to the JL and CL segments, while gene segments in type 2 clusters are all in the same orientation. Although transcripts of type 1 and 3 were readily found in the spleen and head kidney, only minute amounts of type 2 transcripts were seen. The majority of type 3 messages were truncated, suggesting that spliced and full-length transcripts of this isotype probably are present at a low level compared to type 1 transcripts. The uniqueness of the type 3 VLJL sequences suggests that this isotype offers additional diversity to the antigen-binding site of Atlantic salmon immunoglobulins.
通过筛选cDNA文库并分析由组合的3'RACE/5'-锚定PCR产生的DNA,我们在大西洋鲑中分离出三种IgL同种型。其中两种同种型与虹鳟IgL1和L2序列同源,而第三种代表一种先前未表征的鲑科鱼类IgL。新型3型恒定区(CL)与斑点狼鱼c1和黄尾序列同源,而可变区(VL)与沟鲶F类比与任何其他鱼类VL序列更相似。Southern分析表明,所有三种同种型的基因片段都组织成多个簇。此外,3型的VL基因片段相对于连接区(JL)和恒定区(CL)片段以相反的方向排列,而2型簇中的基因片段都以相同的方向排列。虽然在脾脏和头肾中很容易发现1型和3型的转录本,但仅能看到少量的2型转录本。大多数3型信息是截短的,这表明与1型转录本相比,这种同种型的剪接和全长转录本可能以低水平存在。3型VLJL序列的独特性表明,这种同种型为大西洋鲑免疫球蛋白的抗原结合位点提供了额外的多样性。