Subcellular Localization of Signal Peptide Fusion Proteins Expressed in E. coli.
Subcellular Localization of Signal Peptide Fusion Proteins Expressed in E. coli.
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大肠杆菌中表达的信号肽融合蛋白的亚细胞定位。
DOI：
10.1101/pdb.prot102145
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发表时间：
2021
影响因子：
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通讯作者：
Urbatsch,InaL
中科院分区：
文献类型：
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作者：
Kielkopf,ClaraL;Bauer,William;Urbatsch,InaL
对于某些蛋白质在大肠杆菌中的表达，输出至周质空间优于胞质溶胶中的常规表达。输出可以通过将编码序列与编码信号肽的DNA融合(例如，使用pET-22b)来完成，当蛋白质输出到大肠杆菌的内膜和外膜之间的空间时，信号肽被细菌信号肽酶切割。该方案使用渗透压休克从周质中释放多肽。虽然不是定量的，但它应该提供有关信号肽融合蛋白的细胞位置的初步信息。
For expression of some proteins in Escherichia coli, export to the periplasmic space is preferred over conventional expression in the cytosol. Export can be accomplished by fusing the coding sequence to DNA encoding a signal peptide (eg, using pET-22b), which is cleaved by the bacterial signal peptidase as the protein is exported into the space between the inner and outer membranes of E. coli. This protocol uses osmotic shock to release polypeptides from the periplasm. Although not quantitative, it should provide preliminary information on the cellular location of signal peptide fusion proteins.