Ghaffari S H, Lobb C J
Department of Microbiology, University of Mississippi Medical Center, Jackson 39216, USA.
J Immunol. 1997 Jul 1;159(1):250-8.
Earlier studies distinguished two classes of catfish light (L) chain (designated F and G). The cDNA structure and genomic organization of G L chain gene clusters has also been characterized previously. In this study, full length cDNA encoding F L chain was derived using PCR strategies based on the determined amino-terminal protein sequence. The encoded V region is readily delineated into framework regions (FR) and complementarity-determining regions (CDR). Multiple sequence alignments indicate that the F V(L) is closely related to kappa gene families. The F C(L) cannot be generally classified but it is structurally distinct from the C(L) regions of G: the amino acid sequence similarity is <35%. cDNA sequences representing processed sterile F transcripts of different loci were identified. Each sequence begins within the J(L) recombination signal sequence and extends downstream through the I(L)-C(L) segments. Genomic blots hybridized with C(L) probes indicate that there are at least 50 different C(L) segments. Based upon V(L) hybridization studies, different families of V(L) segments appear to be associated with closely related F C(L) segments. In characterized genomic clones, F gene segments are arranged in closely linked clusters with single copies of V(L), J(L), and C(L) segments within each cluster. The V(L) segments are located in opposite transcriptional polarity relative to the J(L) and C(L) segments, which indicates that V(L) segments rearrange by inversion. These combined studies establish that two structurally distinct classes of L chains are present in teleost fish and that both of the L chain classes evolved within a common organizational pattern of clustered segmental genes.
早期研究区分了两类鲶鱼轻(L)链(命名为F和G)。G L链基因簇的cDNA结构和基因组组织先前也已得到表征。在本研究中,基于已确定的氨基末端蛋白质序列,采用PCR策略获得了编码F L链的全长cDNA。所编码的V区很容易被划分为框架区(FR)和互补决定区(CDR)。多序列比对表明,F V(L)与κ基因家族密切相关。F C(L)不能进行一般分类,但在结构上与G的C(L)区不同:氨基酸序列相似性<35%。鉴定出了代表不同基因座经加工的无菌F转录本的cDNA序列。每个序列始于J(L)重组信号序列内,并向下游延伸穿过I(L)-C(L)区段。用C(L)探针杂交的基因组印迹表明至少有50个不同的C(L)区段。基于V(L)杂交研究,不同家族的V(L)区段似乎与密切相关的F C(L)区段相关联。在已表征的基因组克隆中,F基因区段以紧密连锁的簇形式排列,每个簇内有单个拷贝的V(L)、J(L)和C(L)区段。V(L)区段相对于J(L)和C(L)区段处于相反的转录极性,这表明V(L)区段通过倒位进行重排。这些综合研究表明,硬骨鱼中存在两种结构不同的L链类别,并且这两种L链类别都在成簇节段基因的共同组织模式内进化。