• Title/Summary/Keyword: synthetic gene

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연맥의 2배체에서 6배체로의 유전자의 이전에 의한 연구 (Proposal of Gene Transfer Scheme from Diploid (Avena strigosa) to Cultivar Hexaploid (A. sativa) in Oats)

  • 채영암;이종일
    • 한국작물학회지
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    • 제27권3호
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    • pp.243-246
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    • 1982
  • 연맥의 2배체가 가지고 있는 유용한 유전자를 재배종인 6배체에 도입시키는 이론적인 구상을 이미 알려진 사실에 기초하여 논의하였으며 이 계획의 주요 요점은 (1) 서로 다른 배수 수준 사이의 교량 역할을 하는 이질6배체를 얻기 위하여 4배체인 Avena magna를 이용하는 것이다. 이 A. magna는 6배체 재배종인 A. sativa와는 2개의 게놈이 부분적으로 상동성을 가지거나 또는 하나는 공통적이고 나머지 하나는 부분적으로 공통성을 가지고 있기 때문에 AA BB 게놈 형태보다는 이들 사이에 보다 정상적인 접합이 이루어질 수 있기 때문이다. A. strigosa와 A. magna는 교배친화성이며 42% 이상의 임실율을 보이고 있다. (2) A. strigosa의 목적하는 유전자가 A. magna에 도입되어 있는 4배체를 선발하고 (3) 합성 5배체는 $2^n=21$인 모배우자를 생성함으로써 A. sativa와 완전 또는 거의 완전한 접합이 이루어지게 된다. (4) 결과적으로는 마지막 단계에서 $A^{As}$ 게놈에 대하여 이질성이지만 이것을 자식시키면 동질계통을 얻을 수 있게 된다.

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INHIBITORY EFFECT OF OCTYL-PHENOL AND BISPHENOL A ON CALCIUM SIGNALING IN CARDIOMYOCYTE DIFFERENTIATION OF MOUSE EMBRYONIC STEM CELLS

  • J.-H. LEE;Y.-M. YOO;E.-M. JUNG;CH. AHN;E.-B. JEUNG
    • The Korean Journal of Physiology and Pharmacology
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    • 제70권3호
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    • pp.435-442
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    • 2019
  • Endocrine-disrupting chemicals (EDCs) have structures similar to steroid hormones and can interfere with hormone synthesis and normal physiological functions of reproductive organs. For example, sex steroid hormones influence calcium signaling of the cardiac muscle in early embryo development. To confirm the effect of progesterone (P4), octyl-phenol (OP), and bisphenolA(BPA) on early differentiation of mouse embryonic stem cells(mESCs) into cardiomyocytes, mESCs were treated with P4, OP, and BPA two days after attachment and media were replaced every two days. In addition, cells were treated with mifepristone (RU486), a synthetic steroid that has an affinity for progesterone receptor (Pgr), for one day starting on day 11. Beating ratio was decreased with P4, OP, and BPA treatment. The Pgr mRNA level was significantly increased in the P4-, OP- and BPA-treated groups. However, the mRNA level of the calcium channel gene (Trpv2), contraction-related genes (Ryr2, Cam2, and Mylk3) and cardiac development and morphogenesis genes (Rbp4, Ly6e, and Gata4) were significantly decreased in the P4-, OP-, and BPA-treated groups. Interestingly, treatment with RU486 rescued the altered calcium channel gene, contraction-related genes, and cardiac development and morphogenesis genes. P4, OP, and BPA treatments reduced the intracellular calcium level. Taken together, these results indicate that EDCs (OP and BPA) has a structure similar to that of endogenous steroid hormones such as progesterone and estrogen, and OP and BPA act like progesterone to inhibit and disrupt cardiomyocyte differentiation of mESCs.

STK899704 inhibits stemness of cancer stem cells and migration via the FAK-MEK-ERK pathway in HT29 cells

  • Jang, Hui-Ju;Bak, Yesol;Pham, Thu-Huyen;Kwon, Sae-Bom;Kim, Bo-Yeon;Hong, JinTae;Yoon, Do-Young
    • BMB Reports
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    • 제51권11호
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    • pp.596-601
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    • 2018
  • Colon cancer is one of the most lethal and common malignancies worldwide. STK899704, a novel synthetic agent, has been reported to exhibit anticancer effects towards numerous cancer cells. However, the effect of STK899704 on the biological properties of colon cancer, including cancer cell migration and cancer stem cells (CSCs), remains unknown. Here, we examined the inhibitory effect of STK899704 on cell migration and CSC stemness. In the wound healing assay, STK899704 significantly inhibited the motility of colon cancer cells. Furthermore, STK899704 downregulated the mRNA expression levels of the cell migration mediator focal adhesion kinase (FAK). STK899704 also suppressed mitogen-activated protein kinase kinase and extracellular signal-regulated kinase, which are downstream signaling molecules of FAK. Additionally, STK899704 inhibited stemness gene expression and sphere formation in colon cancer stem cells. These results suggest that STK899704 can be used to treat human colon cancer.

Application of sweet and taste modifying genes for development in plants: current status and prospects

  • Akter, Shahina;Huq, Md. Amdadul;Jung, Yu-Jin;Cho, Yong-Gu;Kang, Kwon-Kyoo
    • Journal of Plant Biotechnology
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    • 제43권4호
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    • pp.397-404
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    • 2016
  • Sweet and taste modifying proteins are natural alternatives to synthetic sweeteners and flavor enhancers, and have been used for centuries in different countries. Use of these proteins is limited due to less stability and availability. However, recent advances in biotechnology have enhanced their availability. These include production of sweet and taste modifying proteins in transgenic organisms, and protein engineering to improve their stability. Their increased availability in the food, beverage or medicinal industries as sweeteners and flavor enhancers will reduce the dependence on artificial alternatives. Production of transgenic plants using sweet and taste modifying genes, is an interesting alternative to the extraction of these products from natural source. In this review paper, we briefly describe various sweet and taste modifying proteins (such as thaumatin, monellin, brazzein, curculin and miraculin), their properties, and their application for plant development using biotechnological approaches.

Identification and Characterization of Protein Arginine Methyltransferase 1 in Acanthamoeba castellanii

  • Moon, Eun-Kyung;Kong, Hyun-Hee;Hong, Yeonchul;Lee, Hae-Ahm;Quan, Fu-Shi
    • Parasites, Hosts and Diseases
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    • 제55권2호
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    • pp.109-114
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    • 2017
  • Protein arginine methyltransferase (PRMT) is an important epigenetic regulator in eukaryotic cells. During encystation, an essential process for Acanthamoeba survival, the expression of a lot of genes involved in the encystation process has to be regulated in order to be induced or inhibited. However, the regulation mechanism of these genes is yet unknown. In this study, the full-length 1,059 bp cDNA sequence of Acanthamoeba castellanii PRMT1 (AcPRMT1) was cloned for the first time. The AcPRMT1 protein comprised of 352 amino acids with a SAM-dependent methyltransferase PRMT-type domain. The expression level of AcPRMT1 was highly increased during encystation of A. castellanii. The EGFP-AcPRMT1 fusion protein was distributed over the cytoplasm, but it was mainly localized in the nucleus of Acanthamoeba. Knock down of AcPRMT1 by synthetic siRNA with a complementary sequence failed to form mature cysts. These findings suggested that AcPRMT1 plays a critical role in the regulation of encystation of A. castellanii. The target gene of AcPRMT1 regulation and the detailed mechanisms need to be investigated by further studies.

옥수수 $\alpha$-amylase 유전자의 클로닝 (Cloning of $\alpha$-Amylase Gene from Zea mays)

  • 김용욱;강신혜
    • 한국작물학회지
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    • 제38권3호
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    • pp.275-282
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    • 1993
  • 본 연구는 한국 옥수수의 $\alpha$-amylase의 유전자 클로닝을 주된 목표로 하여 수행되었다. 이를 위하여 여러 식물체의 $\alpha$-amylase 염기서열로 부터 잘 보존된 부분을 참고로 oligonucleotlde probe 및 PCR primer를 설계, 합성하고, 옥수수의 유묘로부터 전체 RNA를 분리하여 northern blot analysis를 통하여 확인한 다음, 이로부터 첫 번째 가닥 cDNA를 만든 후, 여기서 얻은 RNA : DNA hybrid를 주형으로 한 polymerase chain reaction을 통하여 길이 가 약 500bp되 는 PCR 산물을 얻었다. 이를 클로닝하기 위해 pUC19을 클로닝 백터로 사용하여 재조합 플라스미드인 $\ulcorner$pZM$\alpha$'$\lrcorner$를 만들었다. 합성 probe를 이용, Southern blot analysis한 결과, $\ulcorner$pZM$\alpha$'$\lrcorner$가 옥수수 mRNA로 부터 증폭된 DNA의 일부분을 갖고 있음을 확인하였으며, 그 길이는 PCR 산물과 같은 500bp가량 되는 것으로 나타났다.

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Comparison of Ammonia-Oxidizing Bacterial Community Structure in Membrane-Assisted Bioreactors Using PCR-DGGE and FISH

  • Ziembinska, A.;Ciesielski, S.;Gnida, A.;Zabczynski, S.;Surmacz-Gorska, J.;Miksch, K.
    • Journal of Microbiology and Biotechnology
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    • 제22권8호
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    • pp.1035-1043
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    • 2012
  • The ammonia-oxidizing bacterial (AOB) communities in three membrane bioreactors (MBRs) were monitored for 2 months after an acclimation period in order to investigate the influence of sludge age and medium type on AOB changeability and its connection with nitrification effectiveness. One MBR with a sludge age of 4 days was fed with a synthetic medium, whereas the other two with sludge ages of 8 and 32 days were fed with landfill leachate. The research revealed that landfill leachate can be effectively treated in an MBR with a higher sludge age for longer periods of time and that this improvement in performance was correlated with an increase in AOB biodiversity. Interestingly, the medium type has a stronger influence on AOB biocenosis formation than the sludge age.

Recombinant S-Layer Proteins of Lactobacillus brevis Mediating Antibody Adhesion to Calf Intestine Alleviated Neonatal Diarrhea Syndrome

  • Khang, Yong-Ho;Park, Hee-Young;Jeong, Yoo-Seok;Kim, Jung-Ae;Kim, Young-Hwan
    • Journal of Microbiology and Biotechnology
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    • 제19권5호
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    • pp.511-519
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    • 2009
  • A chimeric gene encoding enhanced green fluorescent protein (EGFP) and a S-layer protein from Lactobacillus brevis KCTC3102, and/or two copies of the Fe-binding Z-domain, a synthetic analog of the B-domain of protein A, was constructed and expressed in Escherichia coli BL21(DE3). The S-layer fusion proteins produced in a 500-1 fermentor were likely to be stable in the range of pH 5 to 8 and $0^{\circ}C$ to $40^{\circ}C$. Their adhesive property enabled an easy and rapid immobilization of enzymes or antibodies on solid materials such as plastics, glass, sol-gel films, and intestinal epithelial cells. Owing to their affinity towards intestinal cells and immunoglobulin G, the S-layer fusion proteins enabled the adhesion of antibodies to human epithelial cells. In addition, feeding a mixture of the S-layer fusion proteins and antibodies against neonatal calf diarrhea (coronavirus, rotavirus, Escherichia coli, and Salmonella typhimurium) to Hanwoo calves resulted in 100% prevention of neonatal calf diarrhea syndrome (p<0.01), whereas feeding antibodies only resulted in 56% prevention.

Enhanced Delivery of siRNA Complexes by Sonoporation in Transgenic Rice Cell Suspension Cultures

  • Cheon, Su-Hwan;Lee, Kyoung-Hoon;Kwon, Jun-Young;Choi, Sung-Hun;Song, Mi-Na;Kim, Dong-II
    • Journal of Microbiology and Biotechnology
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    • 제19권8호
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    • pp.781-786
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    • 2009
  • Small interfering synthetic double-stranded RNA (siRNA) was applied to suppress the expression of the human cytotoxic-T-Iymphocyte antigen 4-immunoglobulin (hCTLA4Ig) gene transformed in transgenic rice cell cultures. The sequence of the 21-nucleotide siRNA was deliberately designed and synthesized with overhangs to inactivate the expression of hCTLA4Ig. The chemically synthesized siRNA duplex was combined with polyethyleneimine (PEl) at a mass ratio of 1:10 (0.33 ${\mu}g$ siRNA:3.3 ${\mu}g$ PEl) to produce complexes. The siRNA complexes (siRNA+PEI) were labeled with Cy3 in order to subsequently confirm the delivery by fluorescent microscopy. In addition, the cells were treated with sonoporation at 40 kHz and 419W for 90 s to improve the delivery. The siRNA complexes alone inhibited the expression of hCTLA4Ig to 45% compared with control. The siRNA complexes delivered with sonoporation downregulated the production of hCTLA4Ig to 73%. Therefore, we concluded that the delivery of siRNA complexes into plant cells could be enhanced successfully by sonoporation.

Expression of Sortase, a Transpeptidase for Cell Wall Sorting Reaction, from Staphylococcus aureus ATCC 6538p in Escherichia coli

  • LEE, KI-YOUNG;DONG-SUN SHIN;JUNG-MIN YOON;HEONJOONG KANG;KI-BONG OH
    • Journal of Microbiology and Biotechnology
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    • 제12권3호
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    • pp.530-533
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    • 2002
  • This paper describes the development of an enzymatic assay system for the identification of specific inhibitors of sortase, a transpeptidase that cleaves surface proteins of Cram-positive bacteria, from Staphylococcus aureus ATCC 6538p for antibacterial drug discovery. The coding region of the enzyme was amplified with the exception of the N-terminal membrane anchor sequence, cloned into a vector providing His-Patch-thioredoxin-tag at the N-terminus, expressed in Escherichia coli, and purified by metal chelate affinity chromatography. The enzyme activity was determined by quantifying increased fluorescence intensity upon cleavage of synthetic Dabcyl-QALPETGEE-Edans peptide. The results suggest that the developed in vitro assay system call be used in the search for sortase inhibitors In a short period of time.