Minicells from Highly Genome Reduced Escherichia coli: Cytoplasmic and Surface Expression of Recombinant Proteins and Incorporation in the Minicells,ACS Synthetic Biology

当前位置： X-MOL 学术 › ACS Synth. Biol. › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

Minicells from Highly Genome Reduced Escherichia coli: Cytoplasmic and Surface Expression of Recombinant Proteins and Incorporation in the Minicells
ACS Synthetic Biology ( IF 3.7 ) Pub Date : 2021-09-13 , DOI: 10.1021/acssynbio.1c00375
Hanna Yu ₁ , Andrei V Khokhlatchev ₂ , Claude Chew ₃ , Anuradha Illendula ₂ , Mark Conaway ₄ , Kelly Dryden ₅ , Denicar Lina Nascimento Fabris Maeda ₁ , Vignesh Rajasekaran ₁ , Mark Kester _{2,

6} , Steven L Zeichner _{1,

7}

Affiliation

Minicells, small cells lacking a chromosome, produced by bacteria with mutated min genes, which control cell division septum placement, have many potential uses. Minicells have contributed to basic bacterial physiology studies and can enable new biotechnological applications, including drug delivery and vaccines. Genome-reduced bacteria are another informative area of investigation. Investigators identified that with even almost 30% of the E. coli genome deleted, the bacteria still live. In biotechnology and synthetic biology, genome-reduced bacteria offer certain advantages. With genome-reduced bacteria, more recombinant genes can be placed into genome-reduced chromosomes and fewer cell resources are devoted to purposes apart from biotechnological goals. Here, we show that these two technologies can be combined: min mutants can be made in genome-reduced E. coli. The minC minD mutant genome-reduced E. coli produce minicells that concentrate engineered recombinant proteins within these spherical delivery systems. We expressed recombinant GFP protein in the cytoplasm of genome-reduced bacteria and showed that it is concentrated within the minicells. We also expressed proteins on the surfaces of minicells made from genome-reduced bacteria using a recombinant Gram-negative AIDA-I autotransporter expression cassette. Some autotransporters, like AIDA-I, are concentrated at the bacterial poles, where minicells bud. Recombinant proteins expressed on surfaces of the genome-reduced bacteria are concentrated on the minicells. Minicells made from genome-reduced bacteria may enable useful biotechnological innovations, such as drug delivery vehicles and vaccine immunogens.

中文翻译：

来自高基因组减少大肠杆菌的小细胞：重组蛋白的细胞质和表面表达以及小细胞中的掺入

小细胞是一种缺乏染色体的小细胞，由具有突变的min基因的细菌产生，控制细胞分裂隔膜的位置，具有许多潜在用途。微细胞为基本的细菌生理学研究做出了贡献，并且可以实现新的生物技术应用，包括药物输送和疫苗。基因组减少的细菌是另一个信息丰富的研究领域。调查人员发现，甚至近 30% 的大肠杆菌基因组被删除，细菌仍然活着。在生物技术和合成生物学中，基因组减少的细菌具有一定的优势。使用基因组减少的细菌，可以将更多的重组基因放入基因组减少的染色体中，并且可以将更少的细胞资源用于生物技术目标之外的目的。在这里，我们展示了这两种技术可以结合使用：可以在基因组减少的大肠杆菌中制造最小突变体。minC minD突变基因组减少的大肠杆菌产生在这些球形递送系统中浓缩工程重组蛋白的小细胞。我们在基因组减少的细菌的细胞质中表达重组 GFP 蛋白，并表明它集中在小细胞内。我们还使用重组革兰氏阴性 AIDA-I 自转运蛋白表达盒在由基因组减少的细菌制成的小细胞表面表达蛋白质。一些自转运蛋白，如 AIDA-I，集中在细菌极点，小细胞在那里发芽。在基因组减少的细菌表面上表达的重组蛋白集中在小细胞上。由基因组减少的细菌制成的微细胞可以实现有用的生物技术创新，例如药物输送载体和疫苗免疫原。

更新日期：2021-10-15

点击分享查看原文

点击收藏

公开下载

阅读更多本刊最新论文本刊介绍/投稿指南11