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首页> 美国卫生研究院文献>Proceedings of the National Academy of Sciences of the United States of America >Structural differences in a single gene encoding the V kappa Ser group of light chains explain the existence of two mouse light-chain genetic markers.
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Structural differences in a single gene encoding the V kappa Ser group of light chains explain the existence of two mouse light-chain genetic markers.

机译：编码V kappa Ser轻链组的单个基因的结构差异解释了两个小鼠轻链遗传标记的存在。

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摘要

Two phenotypic markers of mouse immunoglobulin kappa light chains, the IB-peptide marker and the Ef1a isoelectric focusing marker, are expressed by the C58/J, AKR/J, RF/J, and PL/J strains (called expressor strains) but not by BALB/c and most inbred strains. Expression is linked to the kappa light-chain locus and the Lyt-2/Lyt-3 genes on chromosome 6. Light chains bearing these markers belong to a group of variable region kappa chain (V kappa) regions called V kappa Ser, which has a serine amino terminus and a framework 1 region not observed to date among BALB/c light chains. Southern hybridization of genomic DNA with a V kappa Ser-specific cDNA probe has demonstrated a single strongly hybridizing DNA fragment in all strains of mice tested. Characteristic restriction enzyme polymorphisms define the V kappa Ser alleles of expressor (Igk-VSera) and nonexpressor (Igk-VSerb) strains. In the present study, the unrearranged V kappa Ser gene and its flanking regions from an expressor (C.C58) and nonexpressor (BALB/c) strain have been cloned and their nucleotide sequences determined. The C.C58 V kappa Ser gene isolated (the Igk-VSera allele) was shown to code for the two phenotypic markers described. While the nucleotide sequence of the BALB/c coding region (the Igk-VSerb allele) shows 97% identity with the C.C58 gene, single nucleotide substitutions lead to structural changes in the encoded protein which render it IB-negative and Ef1a-negative. These differences alone can explain the failure of strains containing the BALB/c allele to express these kappa-chain phenotypic markers. Also, the BALB/c gene contains a single substitution in a conserved octamer sequence approximately equal to 100 nucleotides upstream of the coding region, which could affect its expression. Finally, the C.C58 allele contains a BAM5/R repetitive DNA element approximately equal to 1200 nucleotides upstream of the coding regions that is not present in BALB/c. This element gives rise to the EcoRI and BamHI restriction enzyme polymorphisms, which distinguish the Igk-VSera and Igk-VSerb alleles.
机译：小鼠免疫球蛋白κ轻链的两个表型标记，IB肽标记和Ef1a等电聚焦标记，由C58 / J，AKR / J，RF / J和PL / J菌株（称为表达菌株）表达，但不表达由BALB / c和大多数自交系组成。表达与6号染色体上的κ轻链基因座和Lyt-2 / Lyt-3基因相关。带有这些标记的轻链属于一组称为V kappa Ser的可变区kappa链（V kappa）区。在BALB / c轻链中迄今尚未观察到的丝氨酸氨基末端和1构架区。基因组DNA与V kappa特异性cDNA探针的Southern杂交已证明在所有测试小鼠品系中都有一个强烈杂交的DNA片段。特征限制酶多态性定义了表达（Igk-VSera）和非表达（Igk-VSerb）菌株的V kappa等位基因。在本研究中，已克隆了未重排的V kappa Ser基因及其来自表达体（C.C58）和非表达体（BALB / c）的侧翼区域，并确定了其核苷酸序列。分离出的C.C58 V kappa Ser基因（Igk-VSera等位基因）显示出所描述的两个表型标记的编码。虽然BALB / c编码区的核苷酸序列（Igk-VSerb等位基因）与C.C58基因具有97％的同一性，但单核苷酸取代会导致编码蛋白的结构变化，从而使其呈IB阴性和Ef1a阴性。 。单独的这些差异可以解释含有BALB / c等位基因的菌株无法表达这些κ链表型标记。同样，BALB / c基因在保守的八聚体序列中包含一个单取代，大约等于编码区上游的100个核苷酸，这可能会影响其表达。最后，C.C58等位基因包含一个BAM5 / R重复DNA元件，大约等于BALB / c中不存在的编码区上游的1200个核苷酸。该元件引起EcoRI和BamHI限制性酶多态性，其区分Igk-VSera和Igk-VSerb等位基因。

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