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Journal : Microbiology Indonesia

 1 
Five Unique Amino Acid Residues of Hemagglutinin (HA) Proteins of Swine Influenza A (H1N1) Detected in 2009 in Jakarta, Indonesia ANDI YASMON; YULIANTY MUHAYAR; VIVI SETIAWATY; BETI ERNAWATI DEWI; BUDIMAN BELA; FERA IBRAHIM
Microbiology Indonesia Vol. 6 No. 2 (2012): June 2012
Publisher : Indonesian Society for microbiology

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (669.535 KB) | DOI: 10.5454/mi.6.2.3

Abstract

Nine HA genes of influenza A (H1N1) viruses originating from swine which were detected in 2009 in Jakarta, Indonesia, were characterized in this study. Nasopharyngeal and/or pharyngeal samples were extracted to obtain viral RNA genomes. Amplification of the HA segment was performed by using the reverse transcription-polymerase chain reaction (RT-PCR), and followed by nested PCR in cases of RT-PCR negative. DNA sequencing was performed by using eight overlapping primers. All the Jakarta strains were closely related to vaccine strain A/California/07/2009. Nine amino acid changes were found in the Jakarta strains, and 5 (P100S, S220T, G239D, R240Q, and I338V) of those were unique to all Jakarta strains with respect to strain A/California/07/2009 used to produce vaccine. An I338V substitution was detected in a cleavage site of HA and no amino acid changes were detected in potential sites for N-linked glycosylation. For seven sites (positions 131, 158, 160, 183, 187, 222, and 227) playing an important role in viral attachment to host receptor, none of the expected amino acid changes was detected; however, a S220T substitution close to amino acid 222 was found in all the Jakarta strains. All amino acid changes potentially affect the pathogenicity of the viruses and the efficacy of strain A/California/07/2009 in neutralizing the Jakarta strains.
Cloning and Expression of HA1 Gene of H1N1 Influenza Virus 2009 Pandemic (H1n1pdm09) Indonesia Strain in the Pichia Pastoris Expression System for the Development of Influenza Vaccine ASRI SULFIANTI; ANDI YASMON; BUDIMAN BELA; FERA IBIRAHIM
Microbiology Indonesia Vol. 9 No. 2 (2015): June 2015
Publisher : Indonesian Society for microbiology

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1209.728 KB) | DOI: 10.5454/mi.9.2.7

Abstract

Among influenza viral proteins, hemagglutinin 1 (HA1) is the target for neutralizing antibodies which inhibit virus binding to receptor of target cells. This protein is widely developed as subunit recombinant vaccine. In this research, we expressed HA1 protein recombinant from DKI271/2011 Indonesian strain in Pichia pastoris. The identity of this protein was confirmed by western blotting using anti-His T ag and mouse specific antibody HA H1N1pdm09. The use of yeast P . pastoris as an alternative strategy to solve the problems which commonly found in influenza vaccine productions. Expression protein in E. coli has been known to have many problems, while mammalian and insect cells requires special skills and relatively high cost. The analysis of HA1 gene sequences showed no mutation in epitope region which recognized by T dan B cells. Further, this recombinant protein can be used as vaccine candidate in influenza vaccine development.Keywords: Hemagglutinin; Pichia pastoris; vaccine; Influenza Virus; H1N1pdm09.
Co-Authors Ade P.R. Simaremare Agusta, Istiqomah Andi Yasmon Andrijono Andrijono Anggraini Alam Anis Anis Ardiana Kusumaningrum Armi Susandi Aroem Naroeni Aroem Naroeni Ascobat, Purwantyastuti ASRI SULFIANTI Beti Ernawati Dewi Cahyani, Hesti Pramesti Christina, Diana Cintera Rahmagiarti Diah Iskandriati Dian Amirulloh Dimas Seto Prasetyo Dondin Sajuthi Elisna Syahruddin Evy Yunihastuti FERA IBIRAHIM Fera Ibrahim Firdaus Sirait Ganjar Noviar Gede Eko Darmono Geraldine, Vanessa Hanifi, Muhammad Hartiyowidi Yuliawuri Hartono, Soni Herna Herna Herna, Herna Hertanto, Robby Ibnu Agus Ariyanto Iin Maemunah Ingrid S. Surono Irawan, Priscilla Felicia Apriliani Jeanne Adiwinata Pawitan Jeanne Elvia Christian Kamila, Etri Dian Ketut Tuti Parwati Lili Indrawati Mardiana Mardiana Maria Lina R. Maria Lina Rosilawati Melinda Remelia Muhayya, Syarifah Raisha Murdani Abdullah Mursinah Mursinah Mursinah Mursinah Ni Ken Ritchie Ni Nengah Dwi Fatmawati Nia Kurniati Nuzli Fahdia Mazfufah Pamungkas, Joko Pauline Phoebe Halim Pramita Gayatri Pratiwi, Ekawati Betty Radiana Dhewayani Antarianto SAHLAN, MUHAMAD Sari, Siska Yuliana Satyapertiwi, Dwiantari SEPTELIA INAWATI WANANDI Silvia Tri Widyaningtyas Silvia Tri Widyaningtyas Silvia Tri Widyaningtyas Silvia Tri Widyaningtyas Silvia Tri Widyaningtyas Silvia Triwidyaningtyas Sofy Meilany Subangkit Subangkit Subiantistha, Tanaya Sugiarta, Gede Yuda SUWIJIYO PRAMONO Triwidyaningtyas, Silvia Utami, Mardhah Sastri VIVI SETIAWATY Widianingtyas, Silvia Widyaningtyas, Silvia Tri Wiseso, Anggoro Yohda, Masafumi YULIANTY MUHAYAR Yuliar Budi Hartanto Yusmaniar Yusmaniar Yuyun SM Soedarmono Yuyun Soedarmono Zakiudin Munasir