在分离的小鼠骨髓瘤细胞核和染色质中进行免疫球蛋白κ轻链基因的体外转录。
Transcription in vitro of immunoglobulin kappa light chain genes in isolated mouse myeloma nuclei and chromatin.
作者信息
Smith M M, Huang R C
出版信息
Proc Natl Acad Sci U S A. 1976 Mar;73(3):775-9. doi: 10.1073/pnas.73.3.775.
Abstract
Messenger RNA sequences for immunoglobulin kappa light chain were synthesized in vitro in isolated mouse myeloma nuclei using bound endogenous RNA polymerase (RNA nucleotidyltransferase; nucleoside triphosphate:RNA nucleotidyltransferase; EC 2.7.7.6) and from isolated myeloma chromatin using exogenous Escherichia coli RNA polymerase. The in vitro RNA was transcribed using 5-mercuriuridine triphosphate and separated from in vivo RNA by chromatography on an agarose sulfhydryl affinity column. Template restriction is retained in vitro since synthesis of kappa chain messenger RNA, As determined by hybridization with complementary DNA, was much more efficient in nuclei and chromatin isolated from myeloma 66.2 tissue culture cells, a kappa-chain-producing cell line, than from MOPC 315 tissue culture cells, a lambda-chain-producing cell line. Transcription of kappa chain messenger RNA was 25 times more efficient in myeloma 66.2 nuclei than in myeloma 66.2 chromatin.
摘要
免疫球蛋白κ轻链的信使核糖核酸序列是在体外，利用结合的内源性RNA聚合酶（RNA核苷酸转移酶；核苷三磷酸：RNA核苷酸转移酶；EC 2.7.7.6）在分离的小鼠骨髓瘤细胞核中合成的，以及利用外源性大肠杆菌RNA聚合酶从分离的骨髓瘤染色质中合成的。体外RNA是使用5-巯基尿苷三磷酸转录的，并通过琼脂糖巯基亲和柱色谱与体内RNA分离。体外保留了模板限制性，因为通过与互补DNA杂交确定，κ链信使核糖核酸的合成在从骨髓瘤66.2组织培养细胞（一种产生κ链的细胞系）分离的细胞核和染色质中比在MOPC 315组织培养细胞（一种产生λ链的细胞系）中效率高得多。κ链信使核糖核酸的转录在骨髓瘤66.2细胞核中的效率比在骨髓瘤66.2染色质中高25倍。
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