Introduction of human gamma 1 immunoglobulin genes into fertilized mouse eggs.
Introduction of human gamma 1 immunoglobulin genes into fertilized mouse eggs.
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将人类 γ1 免疫球蛋白基因引入小鼠受精卵中。
DOI：
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发表时间：
1984
影响因子：
2.7
通讯作者：
T. Kishimoto
中科院分区：
文献类型：
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作者：
K. Yamamura;H. Kikutani;N. Takahashi;T. Taga;S. Akira;K. Kawai;K. Fukuchi;Y. Kumahara;T. Honjo;T. Kishimoto
将重新排列的人类 γ1 免疫球蛋白基因引入小鼠受精卵中。通过将含有VDJ区的pBR322-CESSV(CE-1)的EcoRI和BglII片段与含有γ1恒定区的Ch4A-HIg gamma 1-10的EcoRI和BamHI片段连接来构建噬菌体Ch4A-VCE-γ1。大约 200 个 Ch4A-VCE-gamma 1 基因拷贝被引入小鼠受精卵中。在注射了这些基因的 489 个卵子中，有 319 个存活并被转移到养母的输卵管中。出生了 38 只小鼠，并通过 Southern 印迹杂交筛选了人类 γ1 免疫球蛋白基因的存在。这 38 只小鼠中有 5 只整合了人类 γ 1 免疫球蛋白基因。根据几种限制性内切酶的 Southern 印迹模式判断，每只小鼠中的人类 γ 1 拷贝均未发生删除或重排。人类 γ1 基因存在于几种不同的组织中。迄今为止，所有测试的小鼠都以常染色体显性方式将人类伽马1基因传递给其后代的一部分。通过反向空斑测定或细胞质免疫球蛋白免疫荧光染色分析转基因小鼠脾细胞的免疫球蛋白产生，但无法检测到人γ1链的合成和分泌。在转基因小鼠的肝脏和脾脏中未检测到人γ1免疫球蛋白mRNA。人γ1免疫球蛋白基因的存在似乎对内源性小鼠免疫球蛋白基因的表达没有影响。
A rearranged human gamma 1 immunoglobulin gene was introduced into fertilized mouse eggs. The phage Ch4A-VCE-gamma 1 was constructed by ligating an EcoRI and BglII fragment of pBR322-CESSV(CE-1) containing the VDJ region with an EcoRI and BamHI fragment of Ch4A-HIg gamma 1-10 containing the gamma 1 constant region. About 200 copies of Ch4A-VCE-gamma 1 genes were introduced into fertilized mouse eggs. Of 489 eggs injected with these genes, 319 survived and were transferred to oviducts of foster mothers. Thirtyeight mice were born and were screened for the presence of human gamma 1 immunoglobulin genes by Southern blot hybridization. Five of these 38 mice had integrated human gamma 1 immunoglobulin genes. None of the human gamma 1 copies in each mouse had undergone deletions or rearrangements as judged by the Southern blotting patterns for several restriction enzymes. Human gamma 1 gene was present in several different tissues. All the mice tested so far transmit the human gamma 1 gene to a fraction of their offspring in an autosomal dominant manner. Spleen cells from transgenic mice were analyzed for immunoglobulin production by reverse plaque assay or immunofluorescence staining of cytoplasmic immunoglobulin, but synthesis and secretion of human gamma 1 chains could not be detected. No human gamma 1 immunoglobulin mRNA was detected in the liver and spleen of a transgenic mouse. The presence of the human gamma 1 immunoglobulin gene appeared to have no effect on the expression of endogenous mouse immunoglobulin genes.