Utilizing the Labcyte Echo Liquid Handler and Gateway Gene Assembly in the Generation and Transfection of a PTEN-GFP Construct into a PTEN Null PC-3 Cell Line

The study of protein function relies upon recombinant gene assembly for protein expression and functional assays. This strategy is particularly important when available information regarding function is limited. The vast amount of data generated from genome sequencing projects has paved the way for diverse functional genomic studies that involve cloning, modification and subsequent expression of target genes. This requires flexible and efficient procedures for generating vectors containing gene fusions, gene variants, genetic coding for protein tags and cellular targeting. Furthermore, efficient high throughput cloning and gene assembly procedures are essential for interrogating signaling pathways with multiple gene constructs. The resulting gene constructs can be analyzed in stable transgenic or in transiently expressing biological systems. This application note demonstrates an efficient and robust gene assembly method to generate a fusion protein for functional cellular assays.