• 제목/요약/키워드: DNA density

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Enhanced reutilization value of shrimp-shell waste via fed-batch biodegradation with higher production of reducing sugar, antioxidant, and DNA protective compounds

  • Rashid, Harun Ar;Jung, Hyun Yi;Kim, Joong Kyun
    • Fisheries and Aquatic Sciences
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    • 제21권10호
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    • pp.33.1-33.11
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    • 2018
  • As a process for commercial application, production of reducing sugar, antioxidant, and DNA protective compounds from shrimp-shell powder was investigated in a fed-batch biodegradation using Bacillus cereus EW5. The fed-batch biodegradation was operated in a 5-L bioreactor for 96 h according to three times pulse-feeding strategy. On the basis of the equal working volume (3 L), the fed-batch biodegradation showed a better production of the target compounds than the batch biodegradation, with higher cell density and shortened biodegradation period. The maximum values of the target compounds were 0.297 mg/mL of reducing sugar, 92.35% DPPH radical scavenging activity, 98.16% ABTS radical scavenging activity, and 1.55 reducing power at $A_{700}$, which were approximately 12.1, 3.4, 5.2, and 8.4% enhanced, respectively, compared with those obtained from the batch biodegradation. The fed-batch culture supernatant also showed the enhanced DNA damage inhibition activity than the batch culture supernatant. As a result, the fed-batch biodegradation accompanied by high cell density could produce more useful compounds, enabling an increase in the reutilization value of shrimp-shell waste.

토마토에서 분리된 담배 모자이크 바이러스 외피단백질 유전자의 cDNA 클로닝 및 염기서열 분석 (Complementary DNA Cloning and Nucleotide Sequence Analysis of Coat Protein Gene from TMV Tomato Strain)

  • 이청호;이영기;강신웅;박은경
    • 한국연초학회지
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    • 제18권2호
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    • pp.101-106
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    • 1996
  • Tobacco mosaic virus (TMV) tomato strain was isolated from tomato "Seo-Kwang" in Korea. The virion was purified by density gradient centrifugation, and total viral RNA was isolated from the purified particles. Coat protein (CP) cDNA of the virus was synthesized by RT-PCR, and the purified cDNA fragment was subcloned to pBluescript II SK-. The analysis of nucleotide sequence showed that this cDNA was 693 nucleotides long from the insert of clone p1571 and p1572 which contain complete codons of the viral coat protein gene (474 nucleotides) and 3' untranslated region. The nucleotides of coat protein encoding cDNA of the strain were 6 nucleotides less than that of TMV common strain isolated from tobacco plant in Korea. The CP gene showed 70% maximum homology with that of the common strain in the nucleotide level and 86% maximum homology in amino acid level.cid level.

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저분자량 키토산/유전자 나노콤플렉스 제조 및 이를 이용한 293 세포로의 전달 (DNA Condensation and Delivery in 293 Cells Using Low Molecular Weight Chitosan/gene Nano-complex)

  • 방시원;장양수;김중현;김우식
    • Korean Chemical Engineering Research
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    • 제43권2호
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    • pp.313-317
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    • 2005
  • 양이온성 고분자와 같은 합성 유전자 전달체들은 음이온성을 지닌 plasmid DNA와 쉽게 콤플렉스를 형성하는 경향이 있다. 이에 키토산은 유전자 전달체 시스템으로써 이용되어 질 수 있는 무한한 가능성을 지닌 polysaccharide이다. 저분자량 키토산이 DNA와 결합을 할 수 있는지 확인하기 위하여 전기영동장치를 이용하여 분석하였다. DLS(dynamic laser scattering)와 SEM(scanning electron microscopy)을 이용하여 키토산/DNA 콤플렉스의 크기와 모폴로지를 조사하였다. 또한, 키토산의 분자량과 전하밀도가 콤플렉스의 크기와 결합된 DNA의 양에 어떻게 영향을 주는지 연구를 수행하였다. 저분자량 키토산은 실험과정에서 사용되는 양을 늘려갈수록 84-108%의 세포 생존율을 보임에 따라 그 독성이 무시할 정도가 됨을 확인할 수 있었다. 키토산/DNA 콤플렉스를 이용한 유전자 발현 효율 실험에서는 lipofecamine에 비해서는 낮은 값을 보였지만, naked DNA를 이용한 경우보다는 상대적으로 높은 값을 나타내었다. 키토산의 분자량에 따른 유전자 발현 효율 연구에서는 평균 분자량이 8,517인 키토산을 사용한 경우가 4,078의 분자량을 이용한 실험 결과보다 높은 값을 보였고, 이는 키토산의 전하밀도가 유전자 발현 효율에 영향을 준다는 것을 확인할 수 있었다.

Use of Molecular Replacement to Determine the Phases of Crystal Structure of Taq DNA Polymerase

  • Kim, Young-Soo;Suh, Se-Won
    • BMB Reports
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    • 제29권1호
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    • pp.38-44
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    • 1996
  • Taq DNA polymerase from Thermus aquaticus has been shown to be very useful in the polymerase chain reaction method, which is being used for amplifying DNA. Not only does Taq DNA polymerase have high commercial value commercial value for the polymerase chain reaction application, but it is also important in studying DNA replication, because it is apparently an homologue to E. coli DNA polymerase I, which has long been used for DNA replication study (Lawyer et ai., 1993). The crystal structure determination of Taq DNA polymerase was initiated. An X-ray diffraction pattern breaks down a crystal structure into discrete sine waves in a Fourier series. The original shape of a crystal object in terms of electron density may be represented as the sum of those sine waves with varying amplitudes and phases in three dimensions. The molecular replacement method was initially employed to provide phase information for the structure of Taq DNA polymerase. The rotation search using the program MERLOT resulted in a solution peak with 5.4 r.m.s. PC-refinement of the X-PLOR program verified the result and also optimized the orientation angles. Next, the translation search using the X-PLOR program resulted in a unique solution peak with 7.35 r.m.s. In addition, the translation search indicated $P3_121$ to be the true space group out of two possible ones. The phase information from the molecular replacement was useful in the MIR phasing experiment.

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Immunoliposomes Carrying Plasmid DNA : Preparation and Characterization

  • Kim, Na-Hyung;Park, Hyo-Min;Chung, Soo-Yeon;Go, Eun-Jung;Lee , Hwa-Jeong
    • Archives of Pharmacal Research
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    • 제27권12호
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    • pp.1263-1269
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    • 2004
  • The objective of this study was to characterize immunoliposomes carrying plasmid DNA with optimal encapsulation efficiency and antibody density. Plasmid DNA was encapsulated by the freezing/thawing method into liposomes composed of POPC (1-palmitoyl-2-oleoyl-sn-glycerol- 3-phosphocholine), DDAB (didodecyl dimethyl ammonium bromide), DSPE-PEG 2000 (distearoyl phosphatidyl ethanolamine polyethylene glycol 2000) and DSPE-PEG 2000-maleimide. The liposomes carrying plasmid DNA were extruded through two stacked polycarbonate filters, of different pore size, to control the liposome size. Then, rat IgG molecules were conjugated to the liposomes. The immunoliposomes containing plasmid DNA were separated from the free plasmid DNA and unconjugated IgG by Sepharose CL-4B column chromatography. The DNA amount encapsulated was affected by DDAB (cationic lipid) concentration, the initial amount of plasmid DNA between 10 ${\mu}g$ and 200 ${\mu}g$, the total lipid amount and plasmid DNA size, but not significantly by liposome size. By varying the ratio of DSPE-PEG 2000-maleimide to IgG, the number of IgG molecules per liposome was changed significantly.

Effects of Olanzapine on Gene Expression Changes in MK-801-induced Neurotoxicity Using a High-density DNA Microarray

  • Jo, Jae-Hoon;Kim, Seung-Jun;Yeon, Jong-Pil;Oh, Moon-Ju;Seo, Hye-Myung;Hwang, Seung-Yong;Kim, Sang-Kyum;Kim, Bong-Hee
    • Molecular & Cellular Toxicology
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    • 제3권4호
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    • pp.282-291
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    • 2007
  • Although the etiology of schizophrenia is known to be linked with the disturbance of glutamatergic and dopaminergic neurotransmission, little is known about the relationship between gene expression and the disease process. To identify genes related to abnormalities in glutamatergic and dopaminergic function, we investigated the effects of olanzapine in the changes of mRNA levels in the animal model of schizophrenia, using a high-density DNA microarray. Olanzapine (3.0 mg/kg, i.p.) significantly reduced hyperlocomotive activities, which was induced by MK-801 (1.0 mg/kg, i.p.). We identified that the expression of 719 genes were significantly altered more than two folds in the prefrontal cortex of the rats treated with MK-801. We selected 15 genes out of them by the changes of the expression pattern in the treatment of Olanzapine and/or MK801 for the further confirmation in RT-PCR. The administration of MK-801 increased the expression of 7 genes (NOS3, Hspb1, Hspa1a, CRH, Serpine1, Igfbp6, Snf1lk) and decreased the expression of 1 gene (Aldh1a2), which was attenuated by olanzapine. One gene (Prss12) was up-regulated after olanzapine treatment although it did not show the significant changes after MK-801 treatment. These results showed that antipsychotic drug, such as olanzapine, may alter the gene expression patterns, which were accompanied by MK-801-induced psychosis. Our results also provide us high-density DNA microarray technology could be potential approaches to find the candidate molecules for the therapeutics and also for the early diagnosis of psychiatric diseases.

PCR 기법에 의한 소 수정란의 웅성 특이적 DNA Band 출현과 성 판별에 관한 연구 (Study on Sex Determination and Detection of Male Specific DNA Band in Bovine IVF Embryos Using Polymerase Chain Reaction)

  • 김현종;오성종;김성우;최화식;윤종택;정구민;임경순
    • 한국수정란이식학회지
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    • 제11권3호
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    • pp.283-289
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    • 1996
  • This study was carried out to determine the sex of genomic and embryonic DNA using polymerase chain reaction(PCR). Bovine specific(216bp) and Y chromosome speicific DNA primers(l4lbp) were synthesized and tested for sexing. Bovine embryos used in this study were produced by in vitro fertilization. Few blastomeres for PCR were bisected by nicromanipulator and demi -embryos were cultured in TCM 199 medium containing 0.1% of solcoseryl. The results obtained were as follows; 1. Average optical density of genomic DNA extracted from blood of Hanwoo was 1.79$\pm$ 0.14. 2. 2. The ratio of the demi-embryos developed to blastocyst was 62.1 and 81.9% in morula and blastocyst, respectively. 3. When DNA of 2~4, 5~10 and more than 11 blastomeres was amplified with Y chromosome specific DNA primer by PCR, appreance rate of Y specific DNA band was 16.7, 46.2 and 40.0%, respectively. At least 5 to 10 blastomeres were required to determine the sex of embryos. 4. The rate of demi-embryos developed to blastocyst was 73.3% in TCM 199 medium supplemented with 0.1% solcoceryl. but 55.6% in control.

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DNA Chip 제작을 위한 Microarrayer의 개발 (Development of Microarrayer for DNA Chips)

  • 김석열;정남수;이재성;김상봉
    • 대한기계학회:학술대회논문집
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    • 대한기계학회 2003년도 춘계학술대회
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    • pp.899-904
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    • 2003
  • Microarrayer makes DNA chip and microarray that contain hundreds to thousands of immobilized DNA probes on surface of a microscope slide. This paper shows the development results for a printing type of microarrayer. It realizes a typical, low-cost and efficient microarrayer for generating low density microarray. The microarrayer is developed by using a robot of three-axes perpendicular type. It is composed of a computer-controlled three-axes robot and a pen tip assembly. The key component of the arrayer is the print-head containing the tips to immobilize cDNA, genomic DNA or similar biological material on glass surface. The robot is designed to automatically collect probes from two 96-well plates with up to 32 tips at the same time. To prove the performance of the developed microarrayer, the general water types of inks such as black, blue and red. The inks are distributed at proper positions of 96 well plates and the three color inks are immobilized on the slide glass under the operation procedure. As the result of the test, it can be shown that it has sufficient performance for the production of low integrated DNA chip consisted of 96 spots within 1 $cm^2$ area.

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Channel Capacity Analysis of DNA-based Molecular Communication with Length Encoding Mechanism

  • Xie, Jialin;Liu, Qiang;Yang, Kun;Lin, Lin
    • KSII Transactions on Internet and Information Systems (TIIS)
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    • 제15권8호
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    • pp.2923-2943
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    • 2021
  • The double helix structure of DNA makes it diverse, stable and can store information with high density, and these characteristics are consistent with the requirements of molecular communication for transport carriers. In this paper, a specific structure of molecular communication system based on DNA length coding is proposed. Transmitter (Tx) adopts the multi-layer golden foil design to control the release of DNA molecules of different lengths accurately, and receiver (Rx) adopts an effective and sensitive design of nanopore, and the biological information can be converted to the electric signal at Rx. The effect of some key factors, e.g., the length of time slot, transmission distance, the number of releasing molecules, the priori probability, on channel capacity is demonstrated exhaustively. Moreover, we also compare the transmission capacity of DNA-based molecular communication (DNA-MC) system and concentration-based molecular communication (MC) system under the same parameter setting, and the peak value of capacity of DNA-MC system can achieve 0.08 bps, while the capacity of MC system remains 0.025 bps. The simulation results show that DNA-MC system has obvious advantages over MC system in saving molecular resources and improving transmission stability.

Efficient isolation of sperm with high DNA integrity and stable chromatin packaging by a combination of density-gradient centrifugation and magnetic-activated cell sorting

  • Chi, Hee-Jun;Kwak, Su-Jin;Kim, Seok-Gi;Kim, Youn-Young;Park, Ji-Young;Yoo, Chang-Seok;Park, Il-Hae;Sun, Hong-Gil;Kim, Jae-Won;Lee, Kyeong-Ho
    • Clinical and Experimental Reproductive Medicine
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    • 제43권4호
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    • pp.199-206
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    • 2016
  • Objective: This study was carried out to investigate the correlations of the sperm DNA fragmentation index (DFI) with semen parameters and apoptosis, and to investigate the effects of density-gradient centrifugation (DGC) and magnetic-activated cell sorting (MACS) on reducing the proportion of sperm with DNA fragmentation and protamine deficiency. Methods: Semen analysis and a sperm DNA fragmentation assay were performed to assess the correlations between semen parameters and the DFI in 458 semen samples. Sperm with progressive motility or non-apoptosis were isolated by DGC or MACS, respectively, in 29 normozoospermic semen samples. The effects of DGC or MACS alone and of DGC and MACS combined on reducing the amount of sperm in the sample with DNA fragmentation and protamine deficiency were investigated. Results: The sperm DFI showed a significant correlation (r=-0.347, p< 0.001) with sperm motility and morphology (r=-0.114, p< 0.05) but not with other semen parameters. The DFI ($11.5%{\pm}2.0%$) of semen samples was significantly reduced by DGC ($8.1%{\pm}4.1%$) or MACS alone ($7.4%{\pm}3.9%$) (p< 0.05). The DFI was significantly further reduced by a combination of DGC and MACS ($4.1%{\pm}1.3%$, p< 0.05). Moreover, the combination of DGC and MACS ($1.6%{\pm}1.1%$, p< 0.05) significantly reduced the protamine deficiency rate of semen samples compared to DGC ($4.4%{\pm}3.2%$) or MACS alone ($3.4%{\pm}2.2%$). Conclusion: The combination of DGC and MACS may be an effective method to isolate high-quality sperm with progressive motility, non-apoptosis, high DNA integrity, and low protamine deficiency in clinical use.