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Burcu Sancar Beşen 한국섬유공학회 2019 Fibers and polymers Vol.20 No.12
In the present paper, it was aimed to produce disposable antibacterial textile surfaces with applications of TTOcapsules having different wall materials. For this purpose, tea tree oil (TTO) was encapsulated into β-cyclodextrin (β-CD),polyvinyl alcohol (PVA) and Arabic gum (GA), and the prepared capsules were characterized with scanning electronmicroscope (SEM) and Fourier transform infrared spectroscopy (FTIR) analyses. The capsules, which were proved that theyhad been prepared successfully by the analyses, were applied to the 100 % viscose nonwoven fabric surfaces by paddingmethod at the concentration of 20 % (w/v). The treated fabric samples were characterized through SEM and FTIR analyses,as well as the antibacterial activities of the samples were evaluated against E. coli and S. aureus bacteria. The results indicatedthat the TTO/β-CD, TTO/PVA and TTO/GA capsules could be successfully applied onto textiles and the samples gainedantibacterial activity properties at the different levels depending on the wall materials.
Choi, Jun-Hyuk,Kim, So-Young,Kim, Sook-Kyung,Kemp, Michael G.,Sancar, Aziz American Society for Biochemistry and Molecular Bi 2015 The Journal of biological chemistry Vol.290 No.48
<P>DNA damage by UV and UV-mimetic agents elicits a set of inter-related responses in mammalian cells, including DNA repair, DNA damage checkpoints, and apoptosis. Conventionally, these responses are analyzed separately using different methodologies. Here we describe a unified approach that is capable of quantifying all three responses in parallel using lysates from the same population of cells. We show that a highly sensitive <I>in vivo</I> excision repair assay is capable of detecting nucleotide excision repair of a wide spectrum of DNA lesions (UV damage, chemical carcinogens, and chemotherapeutic drugs) within minutes of damage induction. This method therefore allows for a real-time measure of nucleotide excision repair activity that can be monitored in conjunction with other components of the DNA damage response, including DNA damage checkpoint and apoptotic signaling. This approach therefore provides a convenient and reliable platform for simultaneously examining multiple aspects of the DNA damage response in a single population of cells that can be applied for a diverse array of carcinogenic and chemotherapeutic agents.</P>
Kemp, Michael G.,Gaddameedhi, Shobhan,Choi, Jun-Hyuk,Hu, Jinchuan,Sancar, Aziz American Society for Biochemistry and Molecular Bi 2014 The Journal of biological chemistry Vol.289 No.38
<P>Ultraviolet (UV) photoproducts are removed from genomic DNA by dual incisions in humans in the form of 24- to 32-nucleotide-long oligomers (canonical 30-mers) by the nucleotide excision repair system. How the small, excised, damage-containing DNA oligonucleotides (sedDNAs) are processed in cells following the dual incision event is not known. Here, we demonstrate that sedDNAs are localized to the nucleus in two biochemically distinct forms, which include chromatin-associated, transcription factor II H-bound complexes and more readily solubilized, RPA-bound complexes. Because the nuclear mobility and repair functions of transcription factor II H and RPA are influenced by post-incision gap-filling events, we examined how DNA repair synthesis and DNA ligation affect sedDNA processing. We found that although these gap filling activities are not essential for the dual incision/sedDNA generation event <I>per se</I>, the inhibition of DNA repair synthesis and ligation is associated with a decrease in UV photoproduct removal rate and an accumulation of RPA-sedDNA complexes in the cell. These findings indicate that sedDNA processing and association with repair proteins following the dual incisions may be tightly coordinated with gap filling during nucleotide excision repair <I>in vivo</I>.</P>
Hu, Jinchuan,Choi, Jun-Hyuk,Gaddameedhi, Shobhan,Kemp, Michael G.,Reardon, Joyce T.,Sancar, Aziz American Society for Biochemistry and Molecular Bi 2013 The Journal of biological chemistry Vol.288 No.29
<P>Nucleotide excision repair is the sole mechanism for removing the major UV photoproducts from genomic DNA in human cells. <I>In vitro</I> with human cell-free extract or purified excision repair factors, the damage is removed from naked DNA or nucleosomes in the form of 24- to 32-nucleotide-long oligomers (nominal 30-mer) by dual incisions. Whether the DNA damage is removed from chromatin <I>in vivo</I> in a similar manner and what the fate of the excised oligomer was has not been known previously. Here, we demonstrate that dual incisions occur <I>in vivo</I> identical to the <I>in vitro</I> reaction. Further, we show that transcription-coupled repair, which operates in the absence of the XPC protein, also generates the nominal 30-mer in UV-irradiated XP-C mutant cells. Finally, we report that the excised 30-mer is released from the chromatin in complex with the repair factors TFIIH and XPG. Taken together, our results show the congruence of <I>in vivo</I> and <I>in vitro</I> data on nucleotide excision repair in humans.</P>
Hussein Abdullah Ahmed Ahmed,Selma Onarıcı,Allah Bakhsh,Guray Akdogan,Omer Cem Karakoc,Sancar Fatih Ozcan,Gulsum Aydın,Muhammad Aasım,Levent Unlu,Cengiz Sancak,Samir Naimov,Sebahattin Ozcan 한국식물생명공학회 2017 Plant biotechnology reports Vol.11 No.5
The expression of insecticidal genes must be induced at appropriate time and in sufficient amount to confer protection against targeted pests. However, the increased scientific reports of resistance development in insect pest against insecticidal delta-endotoxins, produced by Bacillus thuringiensis, provide impetus for the development of alternative insect management strategies. The present study was conducted to investigate the importance of targeted expression of a hybrid insecticidal gene (SN19) in potatoes. For this purpose, two plant expression vectors were constructed by cloning hybrid SN19 gene (cry1Badomain I–III and cry1Ia-domain II) under the control of a wound-inducible promoter isolated from Asparagus officinalis (AoPR1) and CaMV 35S promoter, and were transferred to Agrobacterium tumefaciens strain EHA 105. Four potato genotypes (Marabel, Innovator, Tokat 10/1 and Tokat 6/24) were transformed with EHA 105 strain harboring pTF101.1 35S–SN19 and pTF101.1 AoPR1–SN19 constructs. Phosphinothricin (PPT) was used at concentration of 1 mg/l for selection of primary transformants. PCR results showed the presence of both introduced SN19 and bar genes in 43 plants out of total 154 putative transgenics. Expression of SN19 protein in primary transformants was confirmed by Western blot assays. The mechanical wounding of transgenic plants exhibited more accumulated levels of SN19 proteins during post wounding period. Leaf biotoxicity assays with Colorado potato beetle (Coleoptera) and tomato leafminer (Lepidoptera) exhibited 100% mortality of the pests in primary transformants. Based on our mortality results with both constructs, we concluded that the potato transgenic lines exhibited targeted expression of insecticidal gene under the control of AoPR1 promoter upon insect wounding with eliminated toxicity of Cry protein and hence can be further used effectively in potato breeding programme.
Choi, Jun-Hyuk,Gaddameedhi, Shobhan,Kim, So-Young,Hu, Jinchuan,Kemp, Michael G.,Sancar, Aziz Oxford University Press 2014 Nucleic acids research Vol.42 No.4
<P>The nucleotide excision repair pathway removes ultraviolet (UV) photoproducts from the human genome in the form of short oligonucleotides ∼30 nt in length. Because there are limitations to many of the currently available methods for investigating UV photoproduct repair <I>in vivo</I>, we developed a convenient non-radioisotopic method to directly detect DNA excision repair events in human cells. The approach involves extraction of oligonucleotides from UV-irradiated cells, DNA end-labeling with biotin and streptavidin-mediated chemiluminescent detection of the excised UV photoproduct-containing oligonucleotides that are released from the genome during excision repair. Our novel approach is robust, with essentially no signal in the absence of UV or a functional excision repair system. Furthermore, our non-radioisotopic methodology allows for the sensitive detection of excision products within minutes following UV irradiation and does not require additional enrichment steps such as immunoprecipitation. Finally, this technique allows for quantitative measurements of excision repair in human cells. We suggest that the new techniques presented here will be a useful and powerful approach for studying the mechanism of human nucleotide excision repair <I>in vivo</I>.</P>
Emine Anayol,Allah Bakhsh,Omer Cem Karakoc,Selma Onarıcı,Deniz Kom,Muhammad Aasim,Sancar Fatih Ozcan,Surendra Barpete,Saber D. Khabbazi,Burak Onol,Cengiz Sancak,Khalid M. Khawar,Levent Unlu,Sebahattin 한국식물생명공학회 2016 Plant biotechnology reports Vol.10 No.2
Most of the commercialized Bt crops express cry genes under 35S promoter that induces strong gene expression in all plant parts. However, targeted foreign gene expression in plants is esteemed more important as public may be likely to accept ‘less intrusive’ expression of transgene. We developed plant expression constructs harboring cry1Ac gene under control of wound-inducible promoter (AoPR1) to confine Bt gene expression in insect wounding parts of the plants in comparison with cry1Ac gene under the control of 35S promoter. The constructs were used to transform four Turkish cotton cultivars (GSN12, STN-468, Ozbek-100 and Ayhan-107) through Agrobacterium tumefaciens strains GV2260 containing binary vectors p35SAcBAR.101 and AoPR1AcBAR.101 harboring cry1Ac gene under control of 35S and AoPR1, respectively. Phosphinothricin (PPT) was used at concentration of 5 mg L-1 for selection of primary transformants. The primary transformants were analyzed for transgene presence and expression standard molecular techniques. The transformants exhibited appreciable mortality rates against larvae of Spodoptera exigua and S. littoralis. It was found that mechanical wounding of T1 transgenic plants was effective in inducing expression of cry1Ac protein as accumulated levels of cry1Ac protein increased during post-wounding period. We conclude that use of woundinducible promoter to drive insecticidal gene(s) can be regarded as a valuable insect-resistant management strategy since the promoter activity is limited to insect biting sites of plant. There is no Bt toxin accumulation in unwounded plant organs, seed and crop residues, cotton products and by-products, thus minimizing food and environmental concerns.