信使核糖核酸二级结构对大肠杆菌中免疫球蛋白重链表达的影响。
The influence of messenger RNA secondary structure on expression of an immunoglobulin heavy chain in Escherichia coli.
作者信息
Wood C R, Boss M A, Patel T P, Emtage J S
出版信息
Nucleic Acids Res. 1984 May 11;12(9):3937-50. doi: 10.1093/nar/12.9.3937.
Abstract
A gene for murine mu heavy chain immunoglobulin has been inserted into a bacterial expression plasmid containing the Escherichia coli trp promoter and ribosome binding site. A low level expression of mu protein was detected. Secondary structure analysis showed the presence of a hairpin loop burying the mu initiation codon. Alteration of secondary structure at this site by oligonucleotide replacement mutagenesis revealed a correlation between mu expression levels and accessibility of the ribosome binding site. Abolition of secondary structure increased mu protein expression over ninety-fold, to a level approximately equal to that of a trpE -mu fusion protein using the native trpE ribosome binding site.
摘要
已将小鼠μ重链免疫球蛋白基因插入含有大肠杆菌色氨酸启动子和核糖体结合位点的细菌表达质粒中。检测到μ蛋白的低水平表达。二级结构分析表明存在一个发夹环，该发夹环掩盖了μ起始密码子。通过寡核苷酸置换诱变改变该位点的二级结构，揭示了μ表达水平与核糖体结合位点可及性之间的相关性。二级结构的消除使μ蛋白表达增加了90多倍，达到与使用天然色氨酸E核糖体结合位点的色氨酸E-μ融合蛋白大致相当的水平。
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