Molecular Characterization of a Vector-Based Candidate Antigen Using the 3'-RACE and Genome Walking Methods and In Silico Analysis of the Correspondent Protein for Vaccine Design and Development

Abstract

The main objectives of developing vaccines to prevent malaria transmission are malaria control and preventing the reemergence of the disease in endemic regions. Molecular and in silico characterization of a candidate molecule is the first step in the vaccine design process. Determining the sequence and amplification of full-length cDNA copies from the mRNA transcripts is often challenging. The methods in this chapter provide a protocol for the rapid amplification of cDNA ends (RACE) and genome walking. Carboxypeptidase B2 enzyme from A. stephensi (CPBAs-2) was selected as the target molecule and the steps in its characterization and in silico analysis are explained in this chapter.

Keywords: 3′-RACE; 5′-RACE; Carboxypeptidase B2; Genome walking method; Malaria; Molecular characterization; Vaccines that Interrupt Malaria Transmission (VIMTs).