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"Protein Transaktivator transkripsi (Tat) adalah protein regulator HIV-1 berfungsi sebagai aktivator transkripsi genom HIV-1. Varian protein Tat-Eli adalah aktivator transkripsi paling kuat daripada varian lain melalui induksi promotor LTR HVI-1. Kemampuan tersebut digunakan sebagai kontrol positif dalam pengembangan uji infeksitivitas HIV-1 berbasis gene reporter eGFP diregulasi LTR HIV-1. Pengembangan uji infeksivitas tersebut menawarkan waktu deteksi infeksi lebih singkat daripada uji p24 pada uji fenotipik. Penelitian ini bertujuan untuk mengekspresikan protein rekombinan Tat-Eli di sistem ekpresi prokariot dan mempurifikasinya sehingga dapat dijadikan kontrol positif penginduksi promotor. Pada penelitian ini dilakukan pengklonaan gen sintetik Tat-Eli ke vektor pQE80L. Protein rekombinan Tat-Eli dipurifikasi menggunakan Ni-NTA. Pengklonaan ulang gen reporter eGFP disisipkan setelah promotor LTR HIV-1. Aktivitas protein rekombinan Tat-Eli terhadap ekspresi eGFP di sel mamalia dinilai berdasarkan persentase sel pengekspresi eGFP dan intensitas cahaya eGFP.Konstruksi plasmid rekombinan membawa gen Tat-Eli, pQETat, berhasil dibuat, diekspresikan dan dipurifikasi kondisi native. Plasmid pengekspresi eGFP  dengan promoter HIV-1, pLTReGFP berhasil dikonstruksi. Penambahan Tat-Eli rekombinan pada sel mamalia yang ditransfeksi pLTReGFP menunjukkan perbedaan intensitas cahaya eGFP yang bermakna dan paling tinggi dari semua perlakuan. Protein rekombinan Tat-Eli dapat diekspresikan dan dipurifikasi secara optimal dari E.coli. Penambahan protein Tat-Eli pada sel yang ditransfeksi pLTReGFP meningkatkan intensistas cahaya eGFP.

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Transcriptional Transactivator Protein (Tat) is an HIV-1 regulatory protein functioning as an activator of HIV-1 genome transcription. The Tat-Eli protein variant was the most potent transcriptional activator than other variants through the induction of the HVI-1 LTR promoter. This ability was used as a positive control in the development of an HIV-1 infection test based on the eGFP reporter gene regulated by LTR HIV-1. The development of the infectiousness test offers a shorter infection detection time than the p24 test in the phenotypic test. This study aims to express Tat-Eli recombinant protein in the prokaryotic expression system and to purify it so that it can be used as a positive control inducer of the promoter. In this study, synthetic Tat-Eli gene was cloned into the pQE80L vector. Tat-Eli recombinant protein was purified using Ni-NTA. Recloning of the eGFP reporter gene was inserted after the HIV-1 LTR promoter. The activity of Tat-Eli recombinant protein on eGFP expression in mammalian cells was assessed based on the percentage of eGFP-expressing cells and eGFP light intensity. The recombinant plasmid construction carrying the Tat-Eli gene, pQETat was successfully generated, expressed and purified in native conditions. An eGFP-expressing plasmid with HIV-1 promoter, pLTReGFP was successfully constructed. The addition of recombinant Tat-Eli to mammalian cells transfected with pLTReGFP showed a significant difference in eGFP light intensity and was the highest of all treatments. Tat-Eli recombinant protein can be optimally expressed and purified from E. coli. The addition of Tat-Eli protein in pLTReGFP-transfected cells increased eGFP light intensity."

"ABSTRAKUji Western blot masih menjadi gold standard untuk konfirmasi diagnosis infeksi HIV yang memerlukan ketiga protein utama HIV, yaitu env, pol, dan gag. Kekurangan pada uji ini yaitu kemungkinan adanya kontaminasi dengan protein selular manusia serta pembuatannya yang relatif mahal. Selain itu, diversitas HIV-1 yang tinggi menyebabkan uji western blot menjadi kurang sensiftif. Penggunaan antigen rekombinan yang imunodominan dan lestari menjadi alternatif lain. Uji RIBA Recombinant Immunoblot Assay pada penelitian ini menggunakan antigen rekombinan dari keempat subtipe HIV-1 yang dominan di Indonesia, yaitu subtipe CRF01_AE, B, CRF02_AG, dan C. Antigen rekombinan Gag p24 , Pol IDR , Env gp41 IDR diekspresikan pada sistem ekspresi E.coli dan dipurifikasi menggunakan kromatografi Ni-NTA. Antigen rekombinan yang telah dimurnikan dilihat reaktivitasnya terhadap sampel serum pasien dengan HIV-AIDS sebanyak 50 sampel dan non HIV-AIDS 45 sampel. Sebanyak 21 sampel HIV-AIDS dan 3 sampel non HIV-AIDS dilakukan uji menggunakan kit Western blot MP Diagnostics HIV blot 2.2 sebagai perbandingan terhadap uji RIBA yang menggunakan metode Western blot. Hasil perbandingan memperlihatkan hasil uji RIBA memiliki reaktivitas yang lebih baik dibandingkan dengan hasil uji kit MP Diagnostics HIV Blot 2.2 dengan persentase reaktivitas terhadap protein p24 95,2 20/21 , protein Pol 85,7 18/21 , dan protein gp41 100 21/21 . Pada uji RIBA, 5 sampel tidak menunjukkan reaktivitas terhadap antigen rekombinan Pol IDR dan 4 sampel tidak menunjukkan reaktivitas terhadap antigen rekombinan Gag p24 . Seluruh sampel menunjukkan reaktivitasnya terhadap antigen rekombinan Env gp41 IDR . Penelitian mengenai uji RIBA ini dapat dikembangkan untuk uji diagnostik HIV-1 dengan subtipe-subtipe HIV-1 yang banyak ditemukan di Indonesia.

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ABSTRACTern blot test is still the gold standard to confirm the diagnosis of HIV 1 infection. This test requires three core of HIV proteins, i.e., env, pol, and gag. Nevertheless, this test has several disadvantages, mainly in possibility of contamination with human cellular proteins as well as production cost is relatively expensive. In addition, the high diversity of HIV 1 may causes Western blot test to be less sensitive. Another method that can be used to overcome these obstacles is the use of immunodominant region and conserved region of recombinant antigen in the assay, also known as RIBA Recombinant Immunoblot Assay . In this research, recombinant antigens were derived from the four subtypes of HIV 1 that are dominant in Indonesia, which are CRF01 AE subtype, B subtype, CRF02 AG subtype, and C subtype. The recombinant antigens comprises Gag p24 , Pol IDR , Env gp41 IDR . Each of antigens was expressed in E. coli expression system and purified using Ni NTA chromatography. Reactivity test of purified antigen was done against a group consist 50 serum samples with HIV AIDS and 45 serum samples without HIV AIDS. Twenty one samples with HIV AIDS and 3 samples without non HIV AIDS test were done using Western blot kit MP Diagnostics HIV blot 2.2 too as a comparison toward the RIBA test that using Western blot method. The results showed that RIBA test had better reactivity than kit test with reactivity percentage toward p24 95,2 20 21 , Pol 85,7 18 21 , and gp41 100 21 21 . RIBA test results performed 5 samples with negative reactivity toward recombinant antigens Pol IDR and 4 samples with negative reactivity toward recombinant Gag antigen p24 . All the samples had positive reactivity toward recombinan recombinant antigen Env gp41 IDR . As diagnostic kit, this RIBA test shows broad possibility for development in diagnosis HIV 1 infection especially with HIV 1 subtypes that circulate in Indonesia."

"Infeksi HIV-1 merupakan salah satu permasalahan kesehatan di Indonesia yang perlu diteliti untuk mendapatkan strategi intervensi yang sesuai dengan galur virus yang bersirkulasi di Indonesia. Untuk pengembangan kultur galur HIV Indonesia, diperlukan antibodi spesifik terhadap protein awal HIV-1 yaitu Tat dan Rev sebagai marka infeksi HIV pada sel yang terinfeksi HIV-1. Penelitian ini bertujuan untuk mendapatkan plasmid pengekspresi bagian immunodominant protein fusi Tat dan Rev pada sistem ekspresi mamalia agar dapat dimanfaatkan untuk menginduksi pembentukan antibodi spesifik pada hewan coba mamalia. Susunan DNA penyandi protein fusi Tat dan Rev didapat dengan melakukan analisis optimasi kodon penyandi susunan asam amino protein fusi menggunakan piranti lunak GenScript, untuk mendapatkan nilai Codon Adaptation Index CAI > 0.80 pada sistem ekspresi mamalia. Susunan nukleotida yang diperoleh kemudian diberikan tambahan susunan nukleotida situs restriksi pada bagian 5 rsquo; dan 3 rsquo; untuk memudahkan pengklonaan ke dalam vektor ekspresi dan dipesan ke penyedia jasa sintesis asam nukleat. Potongan DNA sisipan TatRev, penyandi protein fusi TatdanRev diperoleh dari penyedia jasa sintesis dalam bentuk terklona dalam plasmid pUC19. Agar dapat terekspresi dalam sistem ekspresi mamalia, DNA sisipan dipotong dengan enzim restriksi KpnI dan HindIII dari plasmid pUC19 dan disubklona ke dalam situs restriksi KpnI dan HindIII pada plasmid pTriEx-4. Analisis restriksi enzim Kpn I dan Hind III plasmid rekombinan yang diperoleh dengan elektroforesis jel agarosa menunjukkan adanya dua pita yang bermigrasi sesuai ukuran plasmid pTriEx-4 dan DNA sisipan TatRev, yaitu sebesar 5000 pb dan 600 pb.

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HIV 1 infection is one of the health problems in Indonesia that is necessary to be studied in order to obtain intervension strategies that is in accordance with the circulating viral strains in Indonesia. In order to develop culture of Indonesian HIV strainss, specific antibodies towards early proteins of HIV, namely Tat and Rev, that are markers of HIV infection in HIV 1 infected cells. This study is aimed at obtaining a plasmid for expression of immunodominant region of Tat and Rev fusion protein in mammalian expression system to be used for stimulation of specific antibody formation in mammals as experimental animal. Nucleotide sequence of DNA encoding the Tat and Rev fusion protein was obtained by codon optimization analysis of the amino acid sequence of the fusion protein using the GenScript software, to obtain a Codon Adaptation Index CAI 0.80 for mammalian expression system. The nucleotide sequence that is obtained was then added with recognition sequences for enzymatic restriction at the 5 rsquo and 3 rsquo end to facilitate cloning into expression vector, and requested to a service provider for nucleic acid synthesis. The TatRev insert DNA, encoding the fusion protein of Tat and Rev was obtained from the nucleic acid synthesis service provider in the formed of cloned DNA in the plasmid pUC19. For expression in mammalian expression system, the insert DNA was restricted using restriction enzymes KpnI and HindIII from the pUC19 plasmid and subcloned into the KpnI and HindIII restriction sites in plasmid pTriEx 4. Agarose gel electrophoresis analysis of KpnI and HindIII enzymatic restriction of the resulting recombinant plasmid showed the presence of two bands that migrated according to the sizes of the plasmid pTriEx 4 and the TatRev insert DNA, namely 5000 bp and 600 bp."

"Acquired Immunodeficiency Syndrome (AIDS) merupakan penyakit akibat infeksi Human Immunodeficiency Virus (HIV) yang memiliki jumlah penderita tinggi di Indonesia. Salah satu upaya untuk mencegah bertambahnya jumlah penderita AIDS tersebut ialah dengan penggunaan vaksin. Poliprotein Gag merupakan protein penyusun struktur internal HIV yang dapat digunakan sebagai vaksin karena dapat menginduksi respon imun tubuh. Penelitian telah dilakukan untuk mengekspresikan poliprotein Gag HIV-1 subtipe CRF01_AE yang telah diinsersi ke dalam vektor ekspresi pQE-80L. Ekspresi poliprotein tersebut dilakukan di dalam bakteri Escherichia coli BL21 dan E. coli BL21-CodonPlus (CP) dengan induksi Isoprophyl-β-D-thiogalactopyranoside (IPTG). Pendeteksian poliprotein Gag hasil ekspresi dilakukan dengan metode Sodium Dodecyl Sulphate Polyacrilamide Gel Electrophoresis (SDS-PAGE). Setelah poliprotein berhasil dideteksi, poliprotein Gag kemudian dipurifikasi dengan menggunakan metode kromatografi afinitas Ni2+-NTA di bawah kondisi native. Poliprotein Gag HIV-1 subtipe CRF01_AE dapat diekspresikan dalam E. coli BL21 dan E. coli BL21-CP dengan berat molekul sebesar 55,3 kDa.

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Acquired Immunodeficiency Syndrome (AIDS) is a disease caused by infection of Human Immunodeficiency Virus (HIV) which has a high number of people in Indonesia. One of the efforts to prevent the increasing number of AIDS patients is the use of vaccine. Gag polyprotein is a constituent protein of HIV internal structure that can be used as a vaccine because it can induce immune response of the body. Research has been conducted to express the Gag polyprotein HIV-1 subtype CRF01_AE that have been cloned into the pQE-80L expression vector. Polyprotein expression was carried out in Escherichia coli BL21 and E. coli BL21-CodonPlus (CP) with Isoprophyl-β-Dthiogalactopyranoside (IPTG) induction. Detection of Gag polyprotein was performed by Polyacrilamide Sodium Dodecyl Sulphate Gel Electrophoresis (SDS-PAGE) method. After successfully detected, Gag polyprotein then purified using Ni2+-NTA affinity chromatography under native condition. Gag polyprotein HIV-1 subtype CRF01_AE can be expressed in E. coli BL21 and E. coli BL21-CP with molecular weight is 55,3 kDa."

"Human Immunodeficiency Virus tipe 1 (HIV-1) merupakan retrovirus penyebab penyakit mematikan AIDS (Acquired Immunodeficiency Syndrome). Virusnya yang cepat bermutasi menyebabkan belum adanya obat yang dapat menyembuhkan penyakit ini secara total. Salah satu target enzim yang dapat diinhibisi untuk menghambat replikasi virus ini adalah protease HIV-1. Inhibisi pada enzim ini menyebabkan hambatan pemotongan protein pada tahap pematangan virus. Beberapa senyawa xanton dari tanaman Garcinia mangostana Linn. yakni α-, β-, dan -mangostin menunjukkan aktivitas inhibisi pada enzim ini. Strukturnya yang berupa nonpeptida memungkinan adanya mekanisme berbeda dari inhibitor lain. Tujuan penelitian ini adalah untuk mengamati model pengikatan dari analog mangostin tersebut. Metode yang digunakan dalam penelitian ini adalah penambatan molekuler secara in silico. Hasil menunjukkan bahwa terdapat dua model pengikatan dengan afinitas yang lebih tinggi pada sisi aktif kantung hidrofobik (ΔG AutoDock 4 = (-9,64)-(-9.89) kkal/mol; ΔG AutoDock Vina = (-8,7)-(-9,4) kkal/mol) dan pada sisi permukaan luar yang masih menunjukkan afinitas ikatan yang baik (ΔG AutoDock 4 = (-5,85)-(-6,06) kkal/mol; ΔG AutoDock Vina = (- 5,3)-(-5,9) kkal/mol).

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Human Immunodeficiency Virus type 1 (HIV-1) is a retroviral virus that cause deadly disease, AIDS (Acquired Immunodeficiency Syndrome). The virus that mutates so fast cause no drugs available to cure this disease totally yet. One of enzyme target that can be inhibited to block the replication of this virus is HIV-1 protease. Inhibition to this enzyme cause the blocking of protein cleavage in virus maturation process. Several xanthones compound from Garcinia mangostana Linn., α-, β-, dan -mangostin, has shown inhibition activity to this enzyme. The structure, which is non-peptide based, gives possibility to different mechanism than other inhibitor. This research?s aim is to search the binding modes of mangostin analogues. The method used in this research is in silico molecular docking. The result shows that there are two binding modes with higher affinity in hydrophobic pocket active site (ΔG AutoDock 4 = (-9,64)-(-9.89) kcal/mol; ΔG AutoDock Vina = (-8,7)-(-9,4) kcal/mol) and molecular surface site which still shows good affinity (ΔG AutoDock 4 = (-5,85)-(-6,06) kcal/mol; ΔG AutoDock Vina = (-5,3)-(-5,9) kcal/mol)."

"This is the first report of HIV drug resistance in RSUPN Dr. Cipto Mangunkusumo. We tested We reviewed eleven new cases of HIV patients who had virologic failure after 6 months first-line antiretroviral therapy. With the sequencing method, analysis of gene mutations encoded HIV drug resistance. Genotypic resistance results and HIV-1 subtype were interpreted by Stanford DR database. Of ten plasma samples that were successfully amplified and sequenced, all samples were resistant to at least one antiretroviral drug. Genotypic resistance towards the antiretroviral drugs being used was observed in lamivudine (90%), tenofovir (83%), nevirapine (100%) dan efavirenz (100%). It is interesting that no zidovudine resistance were found, including in four patients receiving zidovudine in their HAART. The common NRTI mutations were M184VI and K65R, while NNRTI mutations were Y181CFGVY, K103N, A98AG, E138GQ and G190AGS. No mayor PI mutations were found. Based on these findings, we supports the need for appropriate virology monitoring and HIV drug resistance survey in clinical practice and access to drug options in case of virology failure."

"Tat HIV 1 merupakan suatu protein regulator dari HIV 1 yang mempunyai potensi untuk digunakan sebagai vaksin HIV Protein Tat yang diioslasi dari wanita hamil yang menderita HIV di Gabon disebut juga sebagai Tat Oyi mempunyai potensi untuk dijadikan sebagai vaksin dan sistem penghantaran peptida karena sifatnya yang tidak toksik Protein Tat Oyi didapatkan dari hasil Gen Tat Oyi yang telah diekspresikan Ekspresi protein Tat Oyi dapat dilakukan dengan mengekspresikan gen Tat Oyi ke dalam suatu vektor ekspresi Ekspresi Protein membutuhkan Gen Tat Oyi dalam jumlah banyak dan konsentrasi tinggi Pengklonaan gen sintetik Tat Oyi ini dilakukan dalam suatu vektor klona pBluescript KS II Pengklonaan gen sintetik Tat Oyi dilakukan dengan menggunakan pengklonaan dengan ujung blunt pBluescript KS II yang berfungsi sebagai vektor dipotong terlebih dahulu dengan enzim yang memotong dengan ujung blunt EcoRV Plasmid rekombinan Tat Oyi diperbanyak dalam sel inang E coli TOP10 Hasil yang diperoleh dari pengklonaan ini adalah klona gen Tat Oyi dalam plasmid pBluescript KS II Tujuan dilakukan pengklonaan adalah untuk memperbanyak gen Tat Oyi yang akan dibutuhkan dalam proses ekspresi protein Tat Oyi.

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Tat is an regulatory protein of HIV 1 virus that has potential to be used as HIV vaccine Tat Protein from a strain of HIV 1 isolated from Gabon pregnant women that has AIDS also called as Tat Oyi has the potential to be used as vaccine and delivery peptide due to its non toxic property Tat Oyi protein is derived from expressed Tat Oyi gene Tat Oyi protein is expressed by expressing Tat Oyi gene into a expression vector Protein expression into a expression vector will need a lot of high concentration Tat Oyi gene Cloning of this gene is done by clone it into a cloning vector pBluescript KS II This cloning is done with blunt end cloning pBluescript KS II as a vector is restricted with restriction enzim that cut wiht blunt end EcoRV Recombinant Tat Oyi plasmids is cloned into a host cell E coli TOP10 This study resulting Tat Oyi synthetic gene cloned into a plasmid pBluesript KS II The purpose of cloning Tat Oyi gene is to produce a lot of Tat Oyi that will be needed at Tat Oyi protein expression."

"Tanaman dadap ayam (Erythrina variegata L.) mengandung senyawa yang telah diteliti secara in silico mampu menghambat enzim transkiptase balik RT HIV-1 dengan energi pengikatan (∆G) sebesar -10,43 kcal/mol. Selain itu, ekstrak metanol daun dadap ayam menunjukkan adanya aktivitas penghambatan terhadap enzim RT HIV-1 dengan persen penghambatan 97,64% pada konsentrasi 5 mg/ml dibandingkan dengan standar lamivudin. Dengan latar belakang tersebut, dilakukan penelitian untuk mengisolasi senyawa aktif yang terdapat pada ekstrak metanol daun dadap ayam dan dilakukan uji aktivitas terhadap isolat.Uji aktivitas dilakukan secara in vitro menggunakan enzim RT HIV-1 dengan lamivudin sebagai baku pembanding. Uji in vitro menunjukkan fraksi etil asetat sebagai fraksi teraktif dengan nilai IC50 sebesar 429,28 µg/ml. Dari fraksi etil asetat didapatkan senyawa isolat. Senyawa isolat kemudian dielusidasi strukturnya menggunakan spektrofotometer UV, IR, MS, 1H-NMR, 13C-NMR, dan 2D-NMR dan disimpulkan sebagai senyawa apigenin-7-O-β-D-glukopiranosida. Uji in vitro senyawa isolat dengan metode kolorimetri menggunakan kit enzim transkriptase balik menunjukkan nilai IC50 sebesar 100,59 µg/ml sementara lamivudin menunjukkan nilai IC50 sebesar 128,86 µg/ml.

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Erythrina variegata L. containing compounds that have been studied in silico capable to inhibit the HIV-1 reverse transcriptase enzyme with binding energy of -10,43 kcal/mol. Beside that, Erythrina variegata leaves methanolic extract showed an inhibitory activity against HIV-1 RT enzyme with percent inhibition of 97.64% at concentration of 5 mg/mL compared to lamivudine as standard. Depend on this result, this research was held to isolate the active compound as HIV-1 RT inhibitor which is contained in the Erythrina variegata leaves methanolic extract.Activity assay are conducted in vitro using HIV-1 RT colorimetric assay. This assay showed that an ethyl acetate fraction is the most active fraction with IC50 of 429.28 µg/ml. The isolated compound from ethyl acetate fraction was elucidated by spectroscopic tools including UV, IR spectrophotometry, mass, 1D and 2D NMR spectroscopy. The result concluded that an isolated compund as apigenin-7-O-β-D-glucopyranoside. The isolated compound showed inhibitory activity of HIV-1 RT with an IC50 of 100.59 µg/ml compared to lamivudine which showed an IC50 of 128.86 µg/ml."

"Human Immunodeficiency Virus (HIV) adalah virus yang menyerang sistem kekebalan tubuh dan sistem pertahanan infeksi seseorang sehingga menyebabkan penyakit AIDS. Berbagai upaya telah diarahkan untuk pengembangan terapi antiretroviral yang berfokus pada enzim-enzim HIV tipe 1 dengan menggunakan transkriptase balik, protease, dan integrase sebagai target yang potensial. Saat ini, penemuan obat mulai dilakukan dengan pencarian sumber-sumber alam dari ekosistem laut, yang telah dilaporkan memiliki aktivitas biologis, untuk mengeksplorasi berbagai senyawa bioaktif dari organisme laut. Namun, waktu dan biaya yang diperlukan untuk proses penemuan obat sangat besar. Hambatan ini dapat diatasi dengan mengurangi jumlah sampel melalui penggunaan penapisan in silico. Pada penelitian ini, dilakukan pembuatan pangkalan data senyawa kimia dari Echinodermata dan penapisan virtual senyawa-senyawa tersebut terhadap transkriptase balik. Pangkalan data dibuat dengan cara mengumpulkan data dari hasil pencarian literatur yang dilakukan oleh penulis. Penapisan dilakukan menggunakan piranti lunak AutoDock. Berdasarkan hasil penapisan, didapatkan 13 peringkat senyawa terbaik sebagai inhibitor transkriptase balik HIV-1, yaitu Nobilisidenol B, Ech_005, 17-Deoxyholothurinogenin, 22,25-Oxidoholothurinogenin, Ech_022, Ech_026, Ech_021, Nobilisidenol A, Ech_025, 5α-Cholest-8(14)-ene-3ß,7α-diol, Astropectenol A, Ech_004, dan Phrygiasterol.

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Human Immunodeficiency Virus (HIV) is a virus that targets the immune system and weakens people's surveillance and defence systems against infections and causing AIDS disease. Various researches have been directed to the development of antiretroviral therapy that focuses on enzymes of HIV type 1 using reverse transcriptase, protease, and integrase as a potential target. New trends in drug discovery from natural sources emphasize on investigation of the marine ecosystem which have been reported having biological activities to explore bioactive compounds from marine organisms. However, time and cost required for drug discovery process are immense and at an unacceptable level. These obstacles can be overcome by reducing the number of samples using in silico screening. In this research, database compilation of chemical compounds from Echinoderm and virtual screening of the compounds were done to HIV-1 reverse transcriptase. Database was made by collecting data from any references which were searched by author. HIV-1 reverse transkriptase inhibitors virtual screening was done using AutoDock software. Based on the screening results, top thirteen ranked compound was obtained as hits, they are Nobilisidenol B, Ech_005, 17-Deoxyholothurinogenin, 22,25-Oxidoholothurinogenin, Ech_022, Ech_026, Ech_021, Nobilisidenol A, Ech_025, 5α-Cholest-8(14)-ene-3ß,7α-diol, Astropectenol A, Ech_004, and Phrygiasterol."

"ABSTRAKHuman Immunodeficiency Virus HIV merupakan virus yang menyebabkan Acquired Immune Deficiency Syndrome AIDS. Infeksi HIV dapat bersifat laten. Tahapan infeksi meliputi infeksi primer, diseminasi virus ke organ limfoid, peningkatan ekspresi HIV, timbulnya gejala penyakit, dan kematian. Dalam upaya pengendalian kasus infeksi HIV, maka dibutuhkan uji diagnostik serologi yang sensitif dan spesifik. Diagnosis suatu spesimen diawali dengan uji skrining yang berguna untuk identifikasi presumtif kandungan antibodi di dalam spesimen. Salah satu uji skrining yang umum enzyme linked immunosorbent assay ELISA . Uji ELISA untuk diagnosis infeksi HIV saat ini dilakukan berdasarkan antigen, antara lain p24 yang merupakan bagian protein Gag, serta gp41 dan gp120 yang merupakan bagian protein envelope. Telah dilakukan fusi peptida daerah imunodominan gp41 dari 4 subtipe HIV-1 tetraIDR env dengan BSA dalam upaya pengembangan uji ELISA berbasis antigen rekombinan. Gen BSA-tIDR disisipkan ke dalam vektor ekspresi pQE80L dan pengklonaan berhasil menghasilkan plasmid pQE80-BSA-tIDR. Ekspresi protein rekombinan pada bakteri E.coli dilakukan untuk menghasilkan protein BSA-tIDR dan tIDR . Ekspresi protein berhasil dilakukan pada kondisi suhu 37oC, dan dengan induksi IPTG 1 mM selama 4 jam. Protein BSA-tIDR belum berhasil dipurifikasi dengan metode NiNTA. Uji western blot dilakukan terhadap protein hasil ekspresi BSA-tIDR, hasil purifikasi tIDR, dan BSA saja . Hasil uji western blot dengan serum pasien positif HIV-1 memberikan hasil positif pada protein BSA-tIDR dan tIDR.

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ABSTRACTHuman Immunodeficiency Virus HIV is a virus that causes Acquired Immune Deficiency Syndrome AIDS. HIV infection can be latent. Stages of infection include primary infection, viral dissemination to lymphoid organs, increased HIV expression, onset of symptoms, and death. In order to control the HIV infection, a sensitive and specific serologic diagnostic test is required. The diagnosis of a specimen begins with a screening test useful for presumptive identification of the antibody contained in the specimen. One of the common screening tests is enzyme linked immunosorbent assay ELISA . The current ELISA tests for the diagnosis of HIV infection are based on antigens, including p24 which is part of the Gag protein, and gp41 and gp120 which are part of the envelope protein. The fusion of gp41 immunodominant region peptide of 4 HIV 1 subtypes tetraIDR env with BSA has been done to develope recombinant antigen based ELISA assays. The BSA tIDR gene is inserted into the pQE80L expression vector and the cloning successfully produced pQE80 BSA tIDR plasmid. Expression of recombinant protein in E.coli bacteria was performed to produce BSA tIDR and tIDR proteins. The protein expression was successfully performed at 37 C, with 1 mM IPTG induction for 4 hours. BSA tIDR protein has not been successfully purified by NiNTA method. The western blot test was performed on BSA tIDR expression proteins, purified tIDR, and BSA alone. The western blot test with serum HIV 1 positive patients gave positive results on BSA tIDR and tIDR proteins."