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pJoseph2：一种质粒家族，可用作细菌蛋白表达、转染和 Western blot 的阳性对照。

pJoseph2: a family of plasmids as positive controls for bacterial protein expression, transfections, and western blots.

机构信息

Department of Biology, San Diego State University, San Diego, CA 92182, USA.

Department of Biology, University of New Mexico, Albuquerque, NM 87131, USA.

出版信息

Biotechniques. 2024;76(7):299-309. doi: 10.1080/07366205.2024.2343609. Epub 2024 May 12.

DOI:10.1080/07366205.2024.2343609
PMID:39185782
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11796145/
Abstract

Epitope tagging represents a powerful strategy for expedited identification, isolation, and characterization of proteins in molecular biological studies, including protein-protein interactions. We aimed to improve the reproducibility of epitope-tagged protein expression and detection by developing a range of plasmids as positive controls. The pJoseph2 family of expression plasmids functions in diverse cellular environments and cell types to enable the evaluation of transfection efficiency and antibody staining for epitope detection. The expressed green fluorescent proteins harbor five unique epitope tags, and their efficient expression in , Schneider's line 2 cells, and human SKOV3 and HEK293T cells was demonstrated by fluorescence microscopy and western blotting. The pJoseph2 plasmids provide versatile and valuable positive controls for numerous experimental applications.

摘要

表位标记是一种强大的策略，可用于加速分子生物学研究中蛋白质的鉴定、分离和特征分析，包括蛋白质-蛋白质相互作用。我们旨在通过开发一系列质粒作为阳性对照来提高表位标记蛋白表达和检测的重现性。表达质粒 pJoseph2 家族在不同的细胞环境和细胞类型中发挥作用，能够评估转染效率和抗体染色以检测表位。表达的绿色荧光蛋白带有五个独特的表位标签，通过荧光显微镜和 Western blot 证明其在 Schneider's line 2 细胞以及人 SKOV3 和 HEK293T 细胞中的高效表达。pJoseph2 质粒为众多实验应用提供了多功能且有价值的阳性对照。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d2b3/11796145/52f733ca3eb8/nihms-2048541-f0004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d2b3/11796145/692a354ceea8/nihms-2048541-f0001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d2b3/11796145/f8d5b1fb7551/nihms-2048541-f0002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d2b3/11796145/08e3f778cfa6/nihms-2048541-f0003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d2b3/11796145/52f733ca3eb8/nihms-2048541-f0004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d2b3/11796145/692a354ceea8/nihms-2048541-f0001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d2b3/11796145/f8d5b1fb7551/nihms-2048541-f0002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d2b3/11796145/08e3f778cfa6/nihms-2048541-f0003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d2b3/11796145/52f733ca3eb8/nihms-2048541-f0004.jpg

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