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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Immunoglobulin variable region hypermutation in hybrids derived from a pre-B- and a myeloma cell line.
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Immunoglobulin variable region hypermutation in hybrids derived from a pre-B- and a myeloma cell line.

机译：源自前B细胞和骨髓瘤细胞系的杂种中的免疫球蛋白可变区超突变。

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摘要

Somatic mutation of the variable (V) regions of immunoglobulin genes occurs in vivo at rates that have been estimated to be between 10(-3) and 10(-4) per bp per generation. To study this process in vitro, the 18.81 pre-B-cell line and hybrids derived by fusing 18.81 to the NSO myeloma fusion partner were transfected with a mu heavy-chain construct containing a nonsense mutation in the V region (Vn) or the constant region (Cn). Mutation was quantitated by reversion analysis using the ELISA spot assay to detect single cells secreting IgM. Fluctuation analysis revealed that V-region mutations spontaneously occurred in 18.81 cells at an average rate of 5.8 x 10(-6) per bp per cell generation and in selected 18.81-NSO hybrids at greatly increased rates of 1.6 x 10(-3) to 5.8 x 10(-4) per bp per generation. The Vn construct also reverted frequently in transgenic mice, indicating that it contained sufficient information to mutate at high rates both in vivo and in vitro. Sequence analysis of reverted genes revealed that reversion was due to point mutations. Since the rates and nature of the mutations that are occurring in these transfected genes are similar to those reported in vivo, it should be possible to use this system to identify the cis-acting sequences and trans-acting factors that are responsible for V-region somatic hypermutation.
机译：免疫球蛋白基因可变（V）区的体细胞突变在体内发生的速率估计为每代每bp 10（-3）和10（-4）之间。为了在体外研究该过程，将18.81前B细胞系和通过将18.81融合到NSO骨髓瘤融合伴侣而衍生的杂种，用在V区（Vn）或恒定位点无意义突变的mu重链构建体转染。区域（Cn）。通过使用ELISA斑点检测的反向分析对突变进行定量，以检测分泌IgM的单细胞。波动分析表明，V区突变自发地发生在18.81个细胞中，平均每细胞世代bp的速率为5.8 x 10（-6）/ bp，在选定的18.81-NSO杂种中以1.6 x 10（-3）的速率大大增加。每代每bp 5.8 x 10（-4）。 Vn构建体在转基因小鼠中也频繁回复，表明其包含足够的信息以在体内和体外均以高速率突变。回复基因的序列分析表明，回复是由于点突变引起的。由于在这些转染基因中发生的突变的速率和性质与体内报道的相似，因此应该有可能使用该系统来鉴定负责V区的顺式作用序列和反式作用因子体细胞超突变。

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