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未检测到编码小鼠MOPC 41免疫球蛋白轻链mRNA的基因重复。

No detectable reiteration of genes coding for mouse MOPC 41 immunoglobulin light-chain mRNA.

作者信息

Farace M G, Aellen M F, Briand P A, Faust C H, Vassalli P, Mach B

出版信息

Proc Natl Acad Sci U S A. 1976 Mar;73(3):727-31. doi: 10.1073/pnas.73.3.727.

DOI:10.1073/pnas.73.3.727
PMID:815907
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC335991/
Abstract

RNA fractions rich in immunoglobulin light (L)-chain mRNA were isolated from mouse myeloma MOPC 41 by procedures previously described, and chemically labeled with 125I. These RNA fractions were hybridized with MOPC 41 DNA under conditions of DNA excess. Hybridization conditions were chosen under which the entire sequence of the L-chain mRNA probe, thus including the variable region, remains available for hybridization throughout the reaction. The hybridization (C0t) curve showed double transition kinetics, with one component corresponding to about 250 gene copies and the other to about two to four copies. In contrast, when MOPC 41 L-chain mRNA was further purified as a single band by gel elecptrophoresis in 99% formamide, the hybridization curve showed only a single transition, corresponding to about two to four genes, with the disappearance of the "reiterated" component. That component resulted therefore from contaminating RNA species. The data indicate that no reiteration can be detected by RNase or by hydroxylapatite for the genes corresponding to the entire sequence of MOPC 41 L-chain mRNA, including the untranslated segments, within the limits of detectability of short reiterated segments. It thus appears that there is only one or very few genes corresponding to the 41 L-chain variable region "subgroup" in MOPC 41 DNA. The possibility that the variable genes of plasmocytes might result frm a combination of several nonreiterated germline genes is discussed.

摘要

通过先前描述的方法从小鼠骨髓瘤MOPC 41中分离出富含免疫球蛋白轻（L）链mRNA的RNA组分，并用125I进行化学标记。这些RNA组分在DNA过量的条件下与MOPC 41 DNA杂交。选择杂交条件，使得L链mRNA探针的整个序列，包括可变区，在整个反应过程中都可用于杂交。杂交（C0t）曲线显示出双转变动力学，其中一个组分对应于约250个基因拷贝，另一个对应于约2至4个拷贝。相比之下，当MOPC 41 L链mRNA在99%甲酰胺中通过凝胶电泳进一步纯化为单一泳带时，杂交曲线仅显示单一转变，对应于约2至4个基因，“重复”组分消失。因此，该组分是由污染的RNA种类导致的。数据表明，在短重复片段的可检测范围内，对于与MOPC 41 L链mRNA的整个序列（包括非翻译片段）相对应的基因，用核糖核酸酶或羟基磷灰石检测不到重复现象。因此，在MOPC 41 DNA中，似乎只有一个或极少数与41 L链可变区 “亚组” 相对应的基因。文中讨论了浆细胞可变基因可能由几个非重复种系基因组合产生的可能性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0042/335991/a93730913eb4/pnas00672-0070-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0042/335991/a93730913eb4/pnas00672-0070-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0042/335991/a93730913eb4/pnas00672-0070-a.jpg

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SEDIMENTATION STUDIES OF THE SIZE AND SHAPE OF DNA.DNA大小与形状的沉降研究
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