Immunoglobulin from Antarctic fish species of Rajidae family

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• 作者：Maria Rosaria Coscia ; ^{mr.coscia@ibp.cnr.it} ; Ennio Cocca ^{e.cocca@ibp.cnr.it} ; Stefano Giacomelli ^{s.giacomelli@ibp.cnr.it} ; Fausta Cuccaro ^{f.cuccaro@ibp.cnr.it} ; Umberto Oreste ^{u.oreste@ibp.cnr.it}
• 关键词：Antarctic skate ; Cartilaginous fish immunoglobulin M ; C¦Ì gene diversity ; Extra-cysteines
• 刊名：Marine Genomics
• 出版年：2012
• 出版时间：March 2012
• 年：2012
• 卷：5
• 期：Complete
• 页码：35-41
• 全文大小：484 K

文摘

Immunoglobulins (Ig) of Chondroichthyes have been extensively studied in sharks; in contrast, in skates investigations on Ig remain scarce and fragmentary despite the high occurrence of skates in all of the major oceans of the world. To focus on Rajidae Ig¦Ì, the most abundant heavy chain isotype, we have chosen the Antarctic species Bathyraja eatonii, Bathyraja albomaculata, Bathyraja brachyurops, and Amblyraja georgiana which live at high latitudes in the Southern Ocean, and at very low temperatures. We prepared mRNA from the spleen of individuals of each species and performed RT-PCR experiments using two oligonucleotides designed on the alignment of various elasmobranch Ig¦Ì heavy chain sequences available in GenBank. The PCR products, about 1400-nt long, were cloned and sequenced. Nucleotide sequence identities calculated for the constant region domains ranged from 88.5 % to 97.5 % between species, and from 91.1 % to 99.7 % within species. In a distance tree, including also Raja erinacea sequences, two major branches were obtained, one containing Arhynchobatinae sequences, the other one Rajinae sequences. Four presumptive D gene segments were identified in the region of the VH/D/JH recombination; two different D segments were often found in the same sequence. Moreover, 5-15 genomic fragments of different lengths, carrying the gene locus encoding Ig¦Ì chain were revealed by Southern blotting analysis. B. eatonii amino acid sequences were analyzed for the positional diversity by Shannon entropy analysis, showing CH4 as the most conserved domain, and CH3 as the most variable one. B. eatonii CDR3 region length varied between 11 and 15 amino acid residues; the mean length (13.4 aa) was greater than that of Leucoraja eglanteria sequences (7.7 aa). An alignment of representative sequences of Antarctic species and R. erinacea showed that more cysteine residues not involved in the intradomain disulfide bridges were present in Antarctic species.