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Journal : Medical Journal of Indonesia

The production of SPusp45-MSP-119 gene construct and its recombinant protein in Lactococcus lactis to be used as a malaria vaccine Kusuma, Amino V.A.; Mustopa, Apon Z.; Mustafawi, Wike Z.; Suharsono, Suharsono
Medical Journal of Indonesia Vol 26, No 4 (2017): December
Publisher : Faculty of Medicine Universitas Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (574.4 KB) | DOI: 10.13181/mji.v26i4.2162

Abstract

Background: Merozoite surface protein 1 (MSP-1) is a major protein used by the Plasmodium during red blood cells invasion in malaria. MSP-119, one of MSP-1 is highly conserved, and  it is a potential malaria vaccine candidate because the monoclonal antibodies are capable blocking erythrocyte invasion in vitro. The aim of this study was to produce MSP-119 gene construct and the recombinant protein in Lactococcus lactis.Methods: Usp45-MSP-119, derived from codon optimization and the synthetic gene, was inserted into the pMAT cloning vector. A vector expressing MSP-119 included usp45 has been constructed by the manipulation of recombinant DNA using restriction enzymes. The MSP-119 protein was expressed to 45% ammonium sulfate precipitation and purified using Sephadex-G50 gel filtration chromatography. The expressed protein was characterized by SDS-PAGE and dot blot.Results: usp45-MSP-119 gene was amplified using specific primers and inserted into the multiple cloning sites in the expression vector pNZ8148 with size 3,538 bp as a recombinant vector. The protein of  MSP-119 was successfully expressed in L. lactis with molecular weight of 10.45 kDa. The dot blot was tested in 3 different comparisons between the host cells, non-induced cells, and induced cells with 10 ng/ml nisin. The results showed that 10 ng/ml nisin gave a positive reaction as detected by dot blot assay.Conclusion: This study confirmed that the usp45-MSP-119 gene was successfully inserted into the multiple cloning sites of the pNZ8148 expression vector and the MSP-119 protein expressed in the NICE system of the L. lactis host cell.
Co-Authors AA Sudharmawan, AA Adi Isworo, Adi Adikara, Tatang Santanu Agung Setiabudi Aini, Kuratul Akhmad Solikhin AKIHO YOKOTA Al arif, Mohammad Anam Al Firah, Al Firah ALEX HARTANA Alimuddin, Khasan Ana Quthratun Nada Angga Sugiarto Anik Herminingsih Apon Zaenal Mustopa Arif Rianto Budi Nugroho Aris Tjahjoleksono Arwani Arwani Ashadi Ashar N, Novema Aspariansyah, Aspariansyah Asrul Muhamad Fuad, Asrul Muhamad Astanto Kasno Avellyn Shinthya Sari Aviadi, Dedi Badriah, Liah Bambang Sarwono Bambang Yulianto Budianingsih, Budianingsih Cahyadi, Tedy Agung Caswita Caswita Cici Indriani Dalimunthe damayanti, dini Dani Ramdani, Dani Dessy Adelin Destiana, Destiana Didy Sopandie Diki Muhamad Chaidir Diponegoro, Muhammad Djaka Marwasta Egi, Egi Nuryadin Eko Teguh Paripurno Elfa Cahya Jamiati Endang Surahman Erawati, Erna Faisal Rachman Febriana Dwi Wahyuni Feri, Zefki Okta Ferry Faisal, Ferry Firdaus, Syarifin Fitri Damayanti Fitri Damayanti Fitri Wibowo Fitri, Novi Aryani Gandhi Sutjahjo Gesa Cantya Primatara Gunawan, Heru Hadiarto, Toto Hamim Hamim Hapsari, Agustina Retno Harijani, Nenny Hariyadi, Rahmat Hasan Labiqul Aqil Hermana, Salma Junaina Hidanah, Sri Hutabarat, Madya I, Heriberta Reny Idayati, Tutik Rosita Idha Susanti Iif Rahmat Fauzi, Iif Rahmat Ika Barokah Suryaningsih Ika Mariska IMAM MUSTOFA Irbah, Ariq Nurjannah Ishaq, Rizal Maulana Jayawarsa, A.A. Ketut Kagoya, Tundani Kartawan, Kartawan Karuwal, Ritha Lusian KINYA AKASHI Komul, Marsha Krisdiana Wijayanti Kristanto, Paustinus Edi Kusuma, Amino V.A. Kusumayudha, Sari Bahagiarti Kuswanto, Edi Lulut Dwi Sulistyaningsih Lulut Handayani Maemonah, Maemonah Mahmudah, Uswatun Mardiyono Mardiyono marwoto marwoto Meta, Raharni Rosfiyanti Mochamad Widjanarko Mufti Ali Muhammad Helmi Muhammad Jusuf Muhammad Sirril Wafa Muin, Muhammad Rusli Munasik Munasik Mustafawi, Wike Z. Mustofa, Romy Faisal Mustopa, Apon Z. Mutiara, Frista Mycelia Paradise Nabiila, Alyaa Nainggolan, Adinda Rachel Nasrul, Herni Widiyah Ningrum, Hanis Rachna Noor Faidah, Noor Noorikhsan, Faisal Fadilla Novriyandi Hanif Nugroho, Arif Rianto Budi Nugroho, Arif Riyanto Budi Nur Izzati, Aisyah Nuriyatul Fitriya Nurjanah, Fitri Nuryadin, Egi Oktavianti Oktavianti Palaniappan, Ananda Kumar Pausta Yugianus Pertiwi, Nadira Rasya Polosoro, Aqwin Pramono, Sidik Prasetya, Wandy Murti Prasetyadi, C Prasetyadi, Carolus Prasetyadi, Corolus Pratama, Yudha Agung Pujiastuti, Rr Sri Endang purwoko, devit RADITE TISTAMA RATIH DEWI HASTUTI Rikoh Manogar Siringoringo Rinaldi Rizal Putra, Rinaldi Rizal Rivai, Muhamad Rizal Rochadi Abdulhadi Rohananingsih, Rohananingsih Rojabtiyah, Rojabtiyah Rondhianto Rondhianto S. Hidayat, S. Safari, Neng Ayu Mustika Sahputra, Ngatno Sahroni, Sahroni Sahupala, Sapiah Samuel Agus Triyanto, Samuel Agus Santika, Wulan Puja Saputra, Muhammad Wahyu Saputri, Niken Anggraini Sri Sari Bahagiarti Kusumayudha Sari Bahagiarti Kusumayudha Sari, Avellyn Shintya Sari, Fatihah Fajar Sarmanu, Sarmanu Sarsidi Sastrosumarjo Septiana, Erni Setiahadiwibowo, Ajimas Pascaning Shohifaha , Salsabila Sholihin, Achmad Simanjuntak, Veronica Sirait, Budi Chrismanto Sirojuddin, Wildan Siti Zulaeha, Siti sodiq, alwi mahardhika Soekarta, Rendra Solly Aryza Sri Hidanah Sri Langgeng Ratnasari SRI RAHAYU Sri Sumarni Sugiyarto, Angga Sulistyaningsih, Yohana C Sulistyowati, Dina Indrati Dyah Sunarko Sunarko Supranianondo, Koesnoto Susanto, Abiyyu Naufal Sutarto Sutarto Sutrisno, Sutrisno Suyanta Suyanta Ta'adi, Ta'adi Teuku Tajuddin Tita Damayanti Lestari Titik Sudartinah Titin Handayani Trisnaningrum, Niken Trisnowati, Yuni Dwi Trustinah Trustinah Ulfah Juniarti Siregar Utut Widy Astuti UTUT WIDYASTUTI Utut Widyastuti Suharsono Vita Meylani Wahyu Hidayat Wamiliana Wamiliana Wendry Setiyadi Putranto Wening Enggarini Widya Paramita Lokapirnasari Widyastuti, Arum WIJAYANTO WIJAYANTO Winarso, Widyo Wiwik Misaco Yuniarti YanRiska Venata Sembiring, YanRiska Venata Yasir Arafat, Yasir Yeni Yulistanti Yohana Noradika Maharani Yulianto, Bernawan Yuniati, Ratna YUSMANI PRAYOGO Zainal Muttaqin zulfitra, Zulfitra