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小鼠免疫球蛋白λ链恒定区和可变区基因的定量分析

Quantitation of constant and variable region genes for mouse immunoglobulin lambda chains.

作者信息

Honjo T, Packman S

出版信息

Biochemistry. 1976 Jun 29;15(13):2780-5. doi: 10.1021/bi00658a012.

Abstract

We have synthesized and characterized cDNA complementary to purified mRNA derived from the lambda chain producing myeloma tumor, RPC-20. This cDNA is of sufficinet length to encode the constant region and a major portion of the variable region sequence of the lambda gene. In addition, the expected range of cross-hybridization of this lambda probe has been shown to extend to several different members of the closely related lambda subgroup, as well as to a member of the lambda subgroup represented by MOPC-315. Since there are a minimum of seven known members of the common lambda subgroup in addition to MOPC-315, these sequences, in accordance with the germ line hypothesis, must be represented by a minimum of eight variable region genes. Using the RPC-20 cDNA probe and hybridization kinetic analysis, this sequence was found to be represented as approximately two copies per haploid genome in DNA derived from a variety of k-and lambda-producing tumors and normal tissue. Inasmuch as the cross-hybridization range of the probe has been assessed and a minimum size of the lambda subgroup determined, this observation tends to rule out separate germ line genes corresponding to each individual lambda light chain variant. Certain reservations about these conclusions are discussed.

摘要

我们已经合成并鉴定了与源自产生λ链的骨髓瘤肿瘤RPC - 20的纯化mRNA互补的cDNA。该cDNA长度足以编码λ基因的恒定区和可变区序列的主要部分。此外,已证明该λ探针的预期交叉杂交范围延伸至密切相关的λ亚组的几个不同成员,以及由MOPC - 315代表的λ亚组的一个成员。由于除了MOPC - 315之外,常见λ亚组至少还有七个已知成员,根据种系假说,这些序列必须至少由八个可变区基因代表。使用RPC - 20 cDNA探针和杂交动力学分析,发现在源自各种产生κ和λ的肿瘤及正常组织的DNA中,该序列在每个单倍体基因组中约以两个拷贝存在。鉴于已经评估了探针的交叉杂交范围并确定了λ亚组的最小大小,这一观察结果倾向于排除与每个单独的λ轻链变体相对应的单独种系基因。讨论了关于这些结论的某些保留意见。

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