Analysis of Lymphotoxin Alpha Expression in Human Retina and Generation of Expression Vectors to Functional Characterization of Polymorphisms in the Tumor Necrosis Factor Locus

• Published in: Methods in molecular biology (Clifton, N.J.) Vol. 2248; p. 243
• Main Authors: Usategui-Martín, Ricardo, Rodriguez-Hernández, Irene, González-Sarmiento, Rogelio, Sobas, Eva M, Pastor-Jimeno, Jose Carlos, Pastor-Idoate, Salvador
• Format: Journal Article
• Language: English
• Published: United States 2021
• Subjects: Gene Expression; Gene Order; Genetic Loci; Genetic Vectors - genetics; Humans; Lymphotoxin-alpha - genetics; Lymphotoxin-alpha - metabolism; Multigene Family; Polymorphism, Genetic; Retina - metabolism; Tumor Necrosis Factors - genetics; Tumor Necrosis Factors - metabolism; Expression vectors and pCEFL-Flag; Tumor necrosis factor; Site-directed mutagenesis; LTA; Lymphotoxin alpha; Retina; Polymorphisms; Functional characterization
• ISSN: 1940-6029
• DOI: 10.1007/978-1-0716-1130-2_18

With the evolution of new genomic sequencing technologies an important amount of genomic data has been provided. As a consequence of this, many gene polymorphisms have been shown to be significantly associated with different disorders. Many strategies have been implemented to reveal the role of having more than one allele at a specific locus and their involvement in the illnesses. Site-directed mutagenesis is one of the most common strategies to understand the regulatory regions of genes and the relationship between the protein structure and its function. Here, we describe the analysis of lymphotoxin alpha expression in human retina and the generation of expression vectors to functional characterization of polymorphisms in the tumor necrosis factor locus using pCEFL-Flag expression vector and transfection assays in COS-1 cell line.