Nurul Jadid Mubarakati, Aulanni’am Aulanni’am, Sutiman B. Sumitro, Gatot Ciptadi
The aim of this study is to constract the hZP3 gene and to identify the hZP3 clone by using PCR and sequence analysis. Amino acid sequence will be used to identify conserve region of rec-hZP3 using in silico analysis. The sample were derived from human blood. Blood DNA was isolated by salting-out method and then amplified by PCR with a pair of specific primer. The PCR-product was cloned into vector pET-28a and the pET28a-hZP3 clone was transformed into E. coli BL21 competent cells. The pET28a-hZP3 was confirmed by PCR and DNA sequencing. The PCR product of pET28a-hZP3 clone was single band of 122 bp. These results indicated that the hZP3 gene inserted into pET28a properly. In silico analysis showed that amino acid conserved region SQWSRSASRNRR were homology among various species mammalia and have 3 conserved regions of CR 1 (PIECRYPRQGNVSS,137-150 aa), CR 2 (DVTVGPLIFL,359-368 aa) and CR 3 (SQWSRSASRNRR,341-352 aa). The SQWSRSASRNRR sequence confirmed as an epitope recommended for development woman immunocontraception. © 2015, Sphinx Knowledge House. All rights Reserved.
Department of Biology, Islamic University of Malang, Indonesia; School of Veterinary Medicine, Brawijaya University, Indonesia; Biochemistry Laboratory, Department of Chemistry, Brawijaya University, Indonesia; Departement of Biology, Brawijaya University, Malang, Indonesia; Brawijaya University, Indonesia
