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Pcr Amplification of Ornithine Decarboxylase (ODC) Gene Fragment from Tobacco (Nicotiana tabacum L.) cv. Temanggung Djajanegara, Ira; Pambudi, Sabar; Lestari, Retno; Artanti, Nina
ANNALES BOGORIENSES Vol 9, No 2 (2004): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.1234/11

Abstract

In order Lo create an antisense construct of the gene encoding Ornithine Decarboxylase (ODC) from tobacco (Nicotiana tabacum L.) cv. Temanggung, the target gene must be isolated. In this paper. we present the PCR amplification of a fragment from putative gene encoding ODC from tobacco cv. Temanggung. Leaf genomic DNA was i olated and used as the template for PCR. PCR optimization was done by adjusting the annealing temperature and the cycle number. Verification of the fragment obtained was also done using the second primer pairs .  
DNA Condensation Study of Fully Synthesized Lipopeptide-Based Transfection Agent for Gene Delivery Vehicle Tarwadi, Tarwadi; Rachmawati, Heni; Kartasasmita, Rahmana E.; Pambudi, Sabar; Arbianto, Alfan Danny; Restiani, Dewi Esti; Asyarie, Sukmadjaja
ANNALES BOGORIENSES Vol 22, No 2 (2018): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (445.168 KB) | DOI: 10.14203/ann.bogor.2018.v22.n2.65-74

Abstract

   The main requirement of transfection agent has to condense DNA in nanoparticle size, protect the DNA from nucleases and other degrading enzymes during its transport in cell cytoplasm and nucleus and should not toxic to target cells. In this research, lipopeptide composed of palmitoyl (C-16) and short peptide sequence have been designed fully synthesized and tested to DNA condensation capability and toxicity. The DNA condensation study was performed using EtBr exclusion assay and cytotoxicity determination was carried out by colorimetric MTT assay. It was revealed that lipopeptide-based transfection agent of Pal-CKKHH and Pal-CKKHH-YGRKKRRQRRR-PKKKRKV condensed DNA molecules efficiently. The lipopeptide was less toxic compared to Lipofectamine and Poly-L-Lysine, that shown by 90% of CHO-K1 cells remained viable when they were treated with 4.36 µM Pal-CKKHHYGRKKRRQRRR-PKKKRKV. Meanwhile, there were only ~75% and 80% of CHO-K1 viable cells when it was treated with PLL and Lipofectamine®2000, respectively. Moreover, cell viability of HepG2 was ~ 75% after treated with 2.18 µM of Pal-CKKHH-YGRKKRRQRRR-PKKKRKV and decreased to ~65% when the lipopeptide concentration increased to 8.72 M. In summary, the synthesized lipopeptide condenses DNA molecules efficiently, less toxic than Lipofectamine®2000 and PLL and has possibility to be explored as a non-viral gene delivery vehicle.
Pengaruh Ekstrak Etanol Daun Murbei (Morus Alba L.) dengan Glibenklamid Terhadap Ekspresi Gen CYP3A4 pada Kultur Sel HepG2 Nuralih, Nuralih; Churiyah, Churiyah; Pambudi, Sabar; Tamat, Swasono R.; Meila, Okpri
Pharmacon: Jurnal Farmasi Indonesia Vol 15, No 1 (2018)
Publisher : Universitas Muhammadiyah Surakarta

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.23917/pharmacon.v15i1.5766

Abstract

Mulberry leaf is a traditional herb and predicted has ecdysteronecompound which act as antihyperglicemid. Glybenclamide is a synthetic medicine used to cure diabetes mellitus type 2. The leaf reported as competitive inhibitor of CYP3A4 enzyme, which metabolizingglibenclamide. However, in many cases, combination of herbs with synthesis drugs causes interaction if used at the same time. This research aimed to see interaction of ethanol mulberry leaf extract with glibenclamide through CYP3A4 gene expression in HepG2 cell culture.Sample of mulberry extract, glibenclamide, and combination both sample were tested into cell HepG2 culture. Then RNA were isolated and purification using real time PCR to see the gene CYP3A4 expression. As a result, mulberry extract acts as inhibitor enzyme CYP3A4, while glibenclamide is enzyme substrate.The combination of mulberry and glibenclamide showed increased of expression of CYP3A4 gene, means greater enzyme produced, and lower medicine on blood plasma.