The current study was conducted with the main objective to minimize the length of the E-helix of Hpf in order to assemble into 24-mer for possible expression with heterologous proteins for the development of universal vaccine development. Hpf146, Hpf148, Hpf150, Hpf151N147, Hpf151, Hpf152, Hpf153 genes were amplified by PCR using the sense and antisense primers (Table.1). All Hpf truncations were cloned into pET- D u e t vector between B a m H I a n d X h o I by creating an N-terminal His tag. The pET-Duet vector containing the wild-type Hpf gene was used for amplification of all truncations. The bacteria were grown at 37°C, and the plasmid was purified by using a Gene All plasmid mini kit (General Biosystems, Korea), which was analyzed by agarose gel electrophoresis. The clone obtained was confirmed by sequencing. In order to ensure the desired truncations and mutation, sequence was verified via automated DNA sequencing. Proteins were expressed in E. coli B L 2 1 / D E 3. Study findings delineated that all the three constructs viz. Hpf-151N147, Hpf-151 and Hpf- fullL151A showed part of the protein is assembled indicating that at least 151 residues are necessary for its assembly into a 24-mer. Leu151 and Leu153 are the hydrophobic residues of the E-helix, might control the acute angle between the D and E helices formed at the 4-fold axis. The Hpf-fullL151A mutant did not disrupt the assembly, and the purified mutant protein behaved similar to native protein. Leu151 and Leu153 are the hydrophobic residues of the E-helix, might control the acute angle between the D and E helices formed at the 4-fold axis. The Hpf-fullL151A mutant did not disrupt the assembly, and the purified mutant protein behaved similar to native protein