In Silico Gene Analysis and Oligonucleotide Design for the Construction of Expression Vectors

Recombinant expression of a gene of interest is a convenient method to study the function of the encoded protein. To introduce a gene into an expression vector, it is necessary to analyze the structure of the gene and determine some of its basic features including the exact start and end of the coding region or its exon/intron boundaries. Based on this gene analysis, oligonucleotides are then designed for amplification of the coding region by PCR and insertion into the expression vector. For the design of these oligonucleotides, the coding region of the gene of interest and the choice of expression vector must be considered. Here, a number of basic techniques for gene analysis and oligonucleotide design based on specific features of expression vectors are discussed, as well as different methods for introduction of the amplified gene into the expression vector.