• 제목/요약/키워드: Synthetic biology

검색결과 369건 처리시간 0.035초

Design and Cloning of the Gene for a Novel Insulin Analogue, $(B^{30}$-Homoserine) Human Insulin

  • Nam, Doo-H.;Ko, Jeong-Heon;Lee, Seung-Yup
    • Archives of Pharmacal Research
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    • 제16권4호
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    • pp.271-275
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    • 1993
  • In order to prepare a novel human insulin analogue suhbstituted with homoserine at B$^{30}$ / position, (B$^{30}$ /-homoserine) human insulin, a synthetic gene was designed by linking directly a gene for B chain with that for A chain. This gene was constructed by enzymatic joining of 10 different synthetic oligonucleotides, and then inserted at the polylinker region of pUC19 plasmid. To achieve a high level of gene expression, the gene fusion technique region of pUC19 plasmid. To achieve a high level of gene expression, the gene fusion technique was employed using amino terminal regions of lacZ gene up to Clal or hpal, and either of them has been located under tac promoter. The chemical induction of these fused genes by isopropyl-.betha.-D-thiogalactopyranoside (IPTG) gave a satisfactory level of expression in Escherichia coli harboring the ocnstructed plasmids. It was observed that the fused gene product as a single chain insulin precusor was produced more than 30% of total cell protein of E. coli as a form of inclusion body.

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분열효모 Schizosaccharomyces pombe에서 spThp1 유전자 결실돌연변이의 제조와 특성 조사 (Construction of Schizosaccharomyces pombe spThp1 Null Mutants and its Characterization)

  • 윤진호
    • 미생물학회지
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    • 제42권2호
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    • pp.149-152
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    • 2006
  • 분열효모인 Schizosaccharomyces pombe에서 mRNA의 핵에서 세포질로의 이동에 관여할 것으로 여겨지는 spThp1 유전자의 결실돌연변이주(deletion mutant)를 제조하여 그 특성을 조사하였다. 이배체(diploid) 균주의 한 spThp1 유전자를 결실시킨 후 4분체분석(tetrad analysis)을 수행한 결과, 이 유전자는 생장에 필수적이지 않았다. 또한 결실돌연변이주는 mRNA 수송도 큰 결함을 보이지 않았다. 하지만 spThp1 는 mRNA의 운반체를 암호화하고 있는 spMex67와 합성치사(synthetic lethality)를 보였다. 이 결과는 분열효모의 spThpl도 mRNA의 핵에서 세포질로의 이동에 역할을 하고 있음을 암시한다.

적송수피 색소 성분의 화학적 조성과 섬유 염색성 (Dyeabilities with Various Fabrics and Chemical Composition of Brown Colorants from Pine Bark)

  • 김용숙;배순이
    • 한국의류산업학회지
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    • 제15권1호
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    • pp.138-146
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    • 2013
  • Chemical compositions and biological functions of brown colorants extracted from pine bark(Pinus densiflora) have been studied. Dyeing test using multifiber fabrics with extracted colorants were preliminary carried out. Dyeing conditions and fastness tests of selected fabrics have been also studied. The brown colorants were produced 1.5% concentrations by solvent extraction from milled pine bark using methanol. The colorants were extracted with 80% methanol as best choice by a criteria of solid quantity and dyeability on fabrics. The chemical compositions were identified as mixtures of taxifolin epicatechin and procyanidin by LC/MS analysis. The brown colorants could be dyed not only natural fibers such as cotton, silk and wool but also synthetic fiber as nylon and semi-synthetic fiber as viscose rayon. Maximum K/S values was shown at 400 nm according to different fiber with color appearance of redish brown. Optimum pH and temperature of dyeing conditions was 4 and above $80^{\circ}C$, respectively. The brown colorants had a strong antioxidant activity compared to Butylated hydroxyanisole as standard and weak antimicrobial activity against E. coli. compared to kanamycin. Washing, rubbing, perspiration, dry cleaning and light fastness for cotton, nylon and silk dyed with the brown colorants were carried out by KS K method. Most of color fastness such as washing, rubbing, perspiration, and dry cleaning were represented as 4-5 grade. However, light fastness was reported as 2-3 grade. From this studies, brown colorants produced pine bark have a high potentials for natural dyeing on fabrics with antioxidant activity.

미꾸리 자어에 대한 합성세제의 급성독성 (Acute Toxicity of Synthetic Detergent on the Larvae of Loach, Misgurnus angillicaudatus)

  • 이정열;진평
    • 한국수산과학회지
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    • 제17권2호
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    • pp.139-142
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    • 1984
  • 인공수정으로 부화시킨 미꾸리자어에 대한 합성세제(LAS)의 급성독성 영향을 실험하고 그 결과를 생물검정법에 의하여 조사하였다. 1. LAS의 영향을 시간에 대하여 각 농도별로 보면 6ppm이 120시간, 18 ppm이 96시간, 30 ppm이 48시간, 그리고 38 ppm이 16시간만에 $100\%$의 폐사율 가져왔다. 2. 반수치사농도 ($LC_{50}$)는 48hr-$LC_{50}$이 12.59ppm, 96 hr-$LC_{50}$은 4.00 ppm, 120 hr-$LC_{50}$은 1.02 ppm이었다. 3. 급성독성에 대한 영향한계농도는 $0.37{\sim}0.43ppm$이고 적용계수는 $0.093{\sim}0.108$로 나타났다. 4. 합성세제의 각 농도에 대한 반수치사를 가져오는 시간($LT_{50}$)은 0.2 ppm이 165.1시간, 2ppm은 106.2시간으로 나타난 반면 8 ppm은 60.3시간으로 낮아져 38ppm의 경우는 23.5시간이었다.

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Overexpression of Heat Shock Factor Gene HsfA3 Increases Galactinol Levels and Oxidative Stress Tolerance in Arabidopsis

  • Song, Chieun;Chung, Woo Sik;Lim, Chae Oh
    • Molecules and Cells
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    • 제39권6호
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    • pp.477-483
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    • 2016
  • Heat shock factors (Hsfs) are central regulators of abiotic stress responses, especially heat stress responses, in plants. In the current study, we characterized the activity of the Hsf gene HsfA3 in Arabidopsis under oxidative stress conditions. HsfA3 transcription in seedlings was induced by reactive oxygen species (ROS), exogenous hydrogen peroxide ($H_2O_2$), and an endogenous $H_2O_2$ propagator, 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone (DBMIB). HsfA3-overexpressing transgenic plants exhibited increased oxidative stress tolerance compared to untransformed wild-type plants (WT), as revealed by changes in fresh weight, chlorophyll fluorescence, and ion leakage under light conditions. The expression of several genes encoding galactinol synthase (GolS), a key enzyme in the biosynthesis of raffinose family oligosaccharides (RFOs), which function as antioxidants in plant cells, was induced in HsfA3 overexpressors. In addition, galactinol levels were higher in HsfA3 overexpressors than in WT under unstressed conditions. In transient transactivation assays using Arabidopsis leaf protoplasts, HsfA3 activated the transcription of a reporter gene driven by the GolS1 or GolS2 promoter. Electrophoretic mobility shift assays showed that GolS1 and GolS2 are directly regulated by HsfA3. Taken together, these findings provide evidence that GolS1 and GolS2 are directly regulated by HsfA3 and that GolS enzymes play an important role in improving oxidative stress tolerance by increasing galactinol biosynthesis in Arabidopsis.

Generation of Antibodies Against Rice stripe virus Proteins Based on Recombinant Proteins and Synthetic Polypeptides

  • Lian, Sen;Jonson, Miranda Gilda;Cho, Won-Kyong;Choi, Hong-Soo;Je, Yeon-Ho;Kim, Kook-Hyung
    • The Plant Pathology Journal
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    • 제27권1호
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    • pp.37-43
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    • 2011
  • Rice stripe virus (RSV) is one of serious epidemic pathogens for rice species grown in many Asian countries. Therefore, it is necessary to produce a diagnostic detection kit applicable in fields for RSV detection. In this study, RSV proteins that were derived from recombinant proteins and synthetic polypeptides as antigens were generated and were raised in rabbits for antiserum production. Among seven proteins in RSV, genes that code for NCP and NS3 proteins were cloned and subcloned into vector carrying His-tag protein and were expressed in E. coli. Of two recombinant proteins, only anti-NCP displayed stable hybridization signals in western blot analysis. Alternately, synthetic RSV polypeptides for CP, NCP, NS3 and NSvc4 we also generated and only antibodies against CP and NCP were very effective to detect RSV in both RSV infected rice and weed plants. However, antibodies against NS3 and NSvc4 showed weak specific bands as well as strong non-specific background due to the difference of viral proteins produced in the infected leaves. In summary, the antibodies generated against RSV proteins produced in this study will be useful for various assays such as for RSV diagnostic detection, immunoprecipitation, protein purification, and western blot analysis.

Aspergilli에 기생(寄生)하는 Penicillium rugulosum에 대(對)하여 (Penicillium rugulosum Parasite on Aspergilli)

  • 이배함;채희병;이복권;심성보
    • 한국균학회지
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    • 제4권1호
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    • pp.45-52
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    • 1976
  • 기생미생물(寄生微生物)에 대한 연구(硏究)에서 숙주(宿主)와 기생균(寄生菌)을 분류동정(分類同定)하고 성장과정(成長過程)과 그 처리조직학적(處理組織學的) 해부학적(解剖學的) 구조(構造)를 광학(光學) 및 전자현미경(電子顯微鏡)을 통하여 관찰(觀察)하고, 이종간(二種間)의 상호관계(相互關係)를 구명(究明)한 결과(結果)는 다음과 같다. 1. 숙주(宿主)와 기생균(寄生菌)은 Aspergillus niger와 Penicillium rugulosum으로 각각 동정(同定)되었다. 2. 기생균(寄生菌)은 숙주(宿主)의 Sterigmata에 기생(寄生)하고 있는 것을 관찰(觀察)하였다. 3. 기생상태(寄生狀態)의 진행(進行)에 따라 숙주(宿主)의 세포(細胞) 내용물(內容物)이 소실(消失)되어 간다. 4. 기생균(寄生菌)은 합성배지(合成培地)에서도 배양(培養)되므로 활물기생(活物寄生)이 아니다.

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Molecular cloning of a novel cecropin-like peptide gene from the swallowtail butterfly, Papilio xuthus

  • Kim, Seong-Ryul;Choi, Kwang-Ho;Kim, Sung-Wan;Hwang, Jae-Sam;Goo, Tae-Won;Kim, Iksoo
    • International Journal of Industrial Entomology and Biomaterials
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    • 제31권2호
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    • pp.79-84
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    • 2015
  • A new cecropin-like antimicrobial peptide (Px-CLP) gene was isolated from the immunechallenged larvae of the swallowtail butterfly, Papilio xuthus, by employing annealing control primer (ACP)-based GeneFishing PCR. The full-length cDNA of Px-CLP is 310 nucleotides encoding a 70 amino acid precursor that contains a putative 22-residue signal peptide, a 4-residue propeptide, a presumed 37-residue mature peptide, and an uncommon 7-residue acidic pro-region at the C-terminus. The deduced amino acid sequence of Px-CLP showed significant identities with other Lepidopteran cecropin D type peptides. RT-PCR revealed that the Px-CLP transcript was detected at significant level after injection with bacterial lipopolysaccharide (LPS). The peptides with or without C-terminal acidic sequence region were synthesized on-solid phage and submitted to antibacterial activity assay. The synthetic 37-mer peptide (Px-CLPa), which removed C-terminal acidic sequence region, was showed exclusively antibacterial activity against E. coli ML35; meanwhile, a 44-mer peptide (Px-CLPb) with C-terminal acidic peptide region was not active. This result suggests that Px-CLP is produced as a larger precursor containing a C-terminal pro-region that is subsequently removed by C-terminal modification.

Effect of an Anabolic Steroid, Nandrolone Decanoate, on Aquaporin 1 and 9 Gene Expression in the Rat Epididymis

  • Seo, Hee-Jung;Kang, Hyo-Jin;Choi, In-Ho;Cheon, Yong-Pil;Lee, Ki-Ho
    • Reproductive and Developmental Biology
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    • 제33권1호
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    • pp.55-61
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    • 2009
  • The epididymis in the male reproductive tract is the site where spermatozoa produced from the testis become mature. The epididymis is divided into 4 different segments, initial segment and caput, corpus, and caudal epididymis, depending upon functional and morphological features. Aquaporins (Aqps) are water channel molecules, which are present in the epididymis and play a major role in removal of epididymal water, resulting in creation of microenvironment for sperm maturation and concentration of sperms. Nandrolone decanoate (ND) is a synthetic anabolic-androgenic steroid, which is used to treat clinical diseases and improve physical ability and appearance. Even though it is well determined that the ND causes the male infertility by affecting the testis, little is known the effect of the ND on the epididymis. The present study was focused to examine the effect of ND at different treatment doses and periods on expression of Aqp1 and Aqp9 genes in the epididymis of pubertal rats. Results showed that mRNA expression of Aqp1 and Aqp9 genes among the parts of the epididymis was differentially regulated by ND treatment doses. In addition, treatment periods of ND caused differential expression of Aqp1 and Aqp9 mRNAs among segments of the epididymis. Therefore, it is believed that male infertility induced by ND could be resulted not only from malfunction of the testis but also from aberrant gene expression of Aqp1 and Aqp9 in the epididymis.

A Novel Anti-cancer Agent, SJ-8029, Inhibits Angiogenesis and Induces Apoptosis

  • Yi Eui-Yeun;Jeong Eun-Joo;Song Hyun-Seok;Kang Dong-Wook;Joo Jeong-Ho;Kwon Ho-Seok;Lee Sun-Hwan;Park Si-Kyung;Chung Sun-Gan;Cho Eui-Hwan;Kim Yung-Jin
    • 대한의생명과학회지
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    • 제12권3호
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    • pp.161-170
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    • 2006
  • A new piperazine derivative, 8J-8029, is a synthetic anti-cancer agent which exhibits both microtubule and topoisomerase II inhibiting activities. In this study, we investigated the ability of 8J-8029 for anti-angiogenesis and apoptosis. 8J-8029 decreased the bFGF-induced angiogenesis in the CAM and the mouse Matrigel implants, in vivo. 8J-8029 inhibited the proliferation, migration, invasion, tube fonnation, and expression of MMP-2 in BAECs. In addition, 8J-8029 reduced the cell viability in HepG2 cells, caused the production of fragmented DNA and the morphological changes corresponding to apoptosis. 8J-8029 also elicited the release of cytochrome c and the activation of caspase-3. Taken together, these results suggest 8J-8029 may be a candidate for anti-cancer agent with the ability to inhibit the angiogenesis of endothelial cells and to induce the apoptosis of tumor cells.

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