Villatoro-Hernández Julio, Kuipers Oscar P, Saucedo-Cárdenas Odila, Montes-de-Oca-Luna Roberto
Department of Molecular Genetics, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Groningen, The Netherlands.
Methods Mol Biol. 2012;824:155-65. doi: 10.1007/978-1-61779-433-9_8.
This chapter describes the use of Lactococcus lactis as a safe and efficient cell factory to produce heterologous proteins of medical interest. The relevance of the use of this lactic acid bacterium (LAB) is that it is a noncolonizing, nonpathogenic microorganism that can be delivered in vivo at a mucosal level. The use of strains of L. lactis in clinical trials in humans to alleviate inflammatory bowel diseases has opened up the possibility of using this same LAB to target other diseases.Several crucial aspects are addressed in this chapter, such as the expression of heterologous protein, subcellular compartment into which the heterologous protein is located, and description of a standardized protocol to process samples in cell and cell-free fractions to detect the targeted protein expressed by L. lactis.
本章描述了乳酸乳球菌作为一种安全高效的细胞工厂用于生产具有医学价值的异源蛋白的应用。使用这种乳酸菌(LAB)的意义在于它是一种非定殖、非致病的微生物,可以在粘膜水平进行体内递送。乳酸乳球菌菌株在人类临床试验中用于缓解炎症性肠病,这为使用这种相同的乳酸菌来治疗其他疾病开辟了可能性。本章讨论了几个关键方面,例如异源蛋白的表达、异源蛋白所在的亚细胞区室,以及用于处理细胞和无细胞组分中的样品以检测乳酸乳球菌表达的目标蛋白的标准化方案的描述。