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基于 TMP 耐药基因构建并验证小型化质粒

Construction and functional verification of size-reduced plasmids based on TMP resistance gene .

机构信息

Department of Microbiology and Parasitology, Bengbu Medical College , Bengbu, Anhui, China.

Anhui Key Laboratory of Infection and Immunology, Bengbu Medical College , Bengbu, Anhui, China.

出版信息

Microbiol Spectr. 2023 Dec 12;11(6):e0120623. doi: 10.1128/spectrum.01206-23. Epub 2023 Oct 31.

DOI:10.1128/spectrum.01206-23
PMID:37905802
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10714783/
Abstract

Plasmid size is one of the factors affecting transfection efficacy in most of the molecular genetic research studies. One effective approach for reducing plasmid size is to replace relatively large, conventional antibiotic resistance genes with the short-size gene. The successful construct of a series of based tool plasmids and their functional validation, via comparison with original plasmids, suggest that is a potent drug resistance selection marker. The antibiotic trimethoprim offers convenient usage comparable to that of ampicillin or kanamycin. Additionally, fluorescence analysis has demonstrated the compatibility of TMP with protein expression in various host cells. Based on these findings, TMP- could be an alternative choice for future use in molecular genetic research studies that require miniature plasmids to achieve optimal results.

摘要

质粒大小是影响大多数分子遗传学研究中转染效率的因素之一。一种有效的减小质粒大小的方法是用短尺寸的 基因替换相对较大的常规抗生素抗性基因。通过与原始质粒进行比较，成功构建了一系列基于 的工具质粒，并对其功能进行了验证，这表明 是一种有效的抗生素抗性选择标记。抗生素甲氧苄啶的使用方便性可与氨苄青霉素或卡那霉素相媲美。此外，荧光分析表明 TMP 与各种宿主细胞中的蛋白表达兼容。基于这些发现，TMP- 可以成为未来分子遗传学研究中替代使用微型质粒以获得最佳结果的选择。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/63d8/10714783/fb8652260f53/spectrum.01206-23.f008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/63d8/10714783/baf41c8212ae/spectrum.01206-23.f001.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/63d8/10714783/2be959c47399/spectrum.01206-23.f003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/63d8/10714783/9b937180b3cf/spectrum.01206-23.f004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/63d8/10714783/a039c0727839/spectrum.01206-23.f005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/63d8/10714783/bd2216c3d263/spectrum.01206-23.f006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/63d8/10714783/d6bfc2c41561/spectrum.01206-23.f007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/63d8/10714783/fb8652260f53/spectrum.01206-23.f008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/63d8/10714783/baf41c8212ae/spectrum.01206-23.f001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/63d8/10714783/a07b22eab9d1/spectrum.01206-23.f002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/63d8/10714783/2be959c47399/spectrum.01206-23.f003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/63d8/10714783/9b937180b3cf/spectrum.01206-23.f004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/63d8/10714783/a039c0727839/spectrum.01206-23.f005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/63d8/10714783/bd2216c3d263/spectrum.01206-23.f006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/63d8/10714783/d6bfc2c41561/spectrum.01206-23.f007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/63d8/10714783/fb8652260f53/spectrum.01206-23.f008.jpg

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