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斑点叉尾鮰免疫球蛋白轻链的结构与基因组组织。一种不寻常的节段基因基因组组织模式。

Structure and genomic organization of immunoglobulin light chain in the channel catfish. An unusual genomic organizational pattern of segmental genes.

作者信息

Ghaffari S H, Lobb C J

机构信息

Department of Microbiology, University of Mississippi Medical Center, Jackson 39216-4505.

出版信息

J Immunol. 1993 Dec 15;151(12):6900-12.

PMID:8258698
Abstract

Channel catfish L chain cDNA was obtained through a PCR strategy and used to isolate multiple L chain clones from cDNA and genomic libraries. Sequence analysis of full-length cDNA indicates that the V region is preceded by a leader peptide, and represented by framework and CDR regions. Both VL and CL domains contain the invariant cysteines and tryptophans as well as other phylogenetically conserved L chain residues. The sequence similarity of the catfish L chain with higher vertebrate kappa- and lambda-chains, however, does not readily allow the catfish L chain to be classified. Eight cDNA clones isolated from a cDNA library were shown to represent different processed derivatives of sterile L chain transcripts. These transcripts share a similar upstream sequence region and extend downstream to include a CL or alternatively a JL segment in partial germ-line configuration that has been spliced into a CL. Sequence comparisons indicate that these transcripts represent the product of different L chain loci. Genomic Southern blot analyses with VL and CL probes indicate that there are at least 30 VL segments and at least 15 CL segments. The analysis of 17 genomic L chain clones showed that each hybridized with VL-, JL-, and CL-specific probes. Characterization of the gene segments in three of these clones indicates a previously undescribed pattern of segmental gene organization. Gene segments are found in clusters with VL, JL, and CL segments in each cluster. Within a cluster VL segments reside upstream of single copies of closely linked JL and CL segments. The proximity of VL segments downstream from JL-CL segments suggests that individual clusters may be closely linked. The VL segments are located in opposite transcriptional polarity relative to the JL and CL gene segments, which indicates that VL segments are likely rearranged to JL-CL segments by inversion rather than deletion events.

摘要

通过聚合酶链反应(PCR)策略获得了斑点叉尾鮰轻链cDNA,并用于从cDNA和基因组文库中分离多个轻链克隆。全长cDNA的序列分析表明,V区之前有一个前导肽,由框架区和互补决定区(CDR)代表。VL和CL结构域都包含不变的半胱氨酸和色氨酸以及其他系统发育保守的轻链残基。然而,斑点叉尾鮰轻链与高等脊椎动物κ链和λ链的序列相似性并不容易将斑点叉尾鮰轻链进行分类。从一个cDNA文库中分离出的8个cDNA克隆被证明代表无菌轻链转录本的不同加工衍生物。这些转录本共享一个相似的上游序列区域,并向下游延伸,以部分种系构型包含一个CL或一个替代的JL片段,该片段已剪接到一个CL中。序列比较表明,这些转录本代表不同轻链基因座的产物。用VL和CL探针进行的基因组Southern印迹分析表明,至少有30个VL片段和至少15个CL片段。对17个基因组轻链克隆的分析表明,每个克隆都与VL、JL和CL特异性探针杂交。对其中三个克隆中的基因片段进行表征,发现了一种以前未描述的片段基因组织模式。基因片段成簇存在,每个簇中有VL、JL和CL片段。在一个簇中,VL片段位于紧密相连的JL和CL片段单拷贝的上游。JL-CL片段下游的VL片段的接近性表明,各个簇可能紧密相连。VL片段相对于JL和CL基因片段位于相反的转录极性,这表明VL片段可能通过倒位而非缺失事件重排到JL-CL片段。

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