• Title/Summary/Keyword: CHIMERIC

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Studies on Phenotype of Reproduction and Production of Human Growth Hormone(hGH) with Transgenic Rats I. Changes in Endogenous Grwoth Hormone Secretion and Onset of Puberty in hGH Transgenic Rats (Human 성장호르몬을 도입한 Transgenic Rats의 작출과 번식표현형에 관한 연구 I. mWAP/hGH을 도입한 Rat의 Endogenous GH분비 변화와 성성숙에 미치는 영향)

  • 장규태;김성현;성환후;주학진;박미령;윤창현
    • Korean Journal of Animal Reproduction
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    • v.22 no.2
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    • pp.127-136
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    • 1998
  • A chimeric gene comprising murine whey acidic protein(mWAP) and human growth hormone(hGH) was used to produce transgenic rats express hGH and secrete it into the blood. Two lines of transgenic rats carrying the mWAP/hGH construct were established; High line was characterized by relatively high levels of serum hGH, and low line had relatively low levels. The secretory profiles of rat GH(rGH) as well as hGH, the transgene product, were obtained in transgenic males and females of low line; both hGH and rGH serum levels were flattened with no episodic fluctuations, and the overall mean concentration of rGH was significantly lower than in normal littermates. Although the animals of High line showed an acceles, as assessed by vaginal opening and occurrence of first ovulation, advanced by 7∼8 days in both lines of animals. Accordingly, the body weight at puberty of low line transgenic females was much lower than that of normal littermates, indicating that continuous hGH expression could induce precocious puberty without enhancing the growth rate.

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Expression of Low Temperature Regulated Gene H28 in Solanum tuberosum L.

  • Choi, Kyung-Hwa;Jeon, Jae-Heung;Kim, Hyun-Soon;Joung, Young-Hee;Joung, Hyouk;Yang, Deok-Chun
    • Korean Journal of Plant Resources
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    • v.10 no.4
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    • pp.300-304
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    • 1997
  • Leaf dise explants of Solanum tuberosum cultivar. Desiree and Atlantic, were infected with a Agrobacterium MP90 strain containing chimeric gene construct, consisting of antibiotic and low temperature regulated gene (H28) for transformation. regenerated multiple shoots were selected on a medium containing kanamycin and carbenieillin after exposure to Agrobacterium. Both PCR analysis of NPT Ⅱ, H28 genes and northern blot analysis indicated that the genes coding for the enzyme were successfully integrated into the potato genome and could be expressed in potato plants.

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Up-Regulation of Glutathione Biosynthesis in NIH3T3 Cells Transformed with the ETV6-NTRK3 Gene Fusion

  • Kim, Su-Jung;Kim, Hong-Gyum;Lim, Hye-Won;Park, Eun-Hee;Lim, Chang-Jin
    • Molecules and Cells
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    • v.19 no.1
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    • pp.131-136
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    • 2005
  • The ETV6-NTRK3 gene fusion, first identified in the chromosomal translocation in congenital fibrosarcoma, encodes a chimeric protein tyrosine kinase with potent transforming activity. ETV6-NTRK3-dependent transformation involves the joint action of NTRK3 signaling pathways, and aberrant cell cycle progression resulting from activation of Mek1 and Akt. The level of glutathione (GSH) was found to be markedly increased in ETV6-NTRK3-transformed NIH3T3 cells. The activities of the two GSH biosynthetic enzymes as well as of glutathione peroxidase, together with their mRNAs, were also higher in the transformed cells. The transformed cells were able to grow in the presence of GSH-depleting agents, whereas the control cells were not. L-Buthionine-(S,R)-sulfoximine (BSO) inhibited activation of Mek1 and Akt in the transformed NIH3T3 cells. These observations imply that up-regulation of GSH biosynthesis plays a central role in ETV6-NTRK3-induced transformation.

Development of Leuconostoc sp. Host Vector System

  • Eom, Hyun-Ju;Park, Myeong-Soo;Ji, Geun-Eog;Han, Nam-Soo
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2004.06a
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    • pp.323-327
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    • 2004
  • Leuconostoc citreum CBUE isolated from kimchi proved to harbor a small cryptic plasmid, pNS75. The complete nucleotide sequence of pNS75 was 1,821 bp and had a low G+C content of 39.2%. Computer analysis using DNASIS revealed one open reading frame (ORF), having ATG as putatitive start condon and potentially encoding proteins with molecular mass of 38 kDa. The chimeric plasmid pLeuCM was first constructed wih pNS75, pUC19 and chroamphenicol acetyltransferase (CAT) from Staphylococcus sp.. pLeuCM replicated and expressed chroamphenicol acetyltransferase in Leuconostoc citerum CBNF after transformation. To test the availability of shuttle vector as cloning vehicle of foreign gene, $\alpha$-amylase gene of Streptococcus bovis was cloned and all transformants secreated the $\alpha$-amylase successfully. The result indicates that pLeuCM is a potential shuttle vector for Leuconostoc spp. and lactic acid bacteria.

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Development of a Reporter System for In Vivo Monitoring of γ-Secretase Activity in Drosophila

  • Hong, Young Gi;Roh, Seyun;Paik, Donggi;Jeong, Sangyun
    • Molecules and Cells
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    • v.40 no.1
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    • pp.73-81
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    • 2017
  • The ${\gamma}$-secretase complex represents an evolutionarily conserved family of transmembrane aspartyl proteases that cleave numerous type-I membrane proteins, including the ${\beta}$-amyloid precursor protein (APP) and the receptor Notch. All known rare mutations in APP and the ${\gamma}$-secretase catalytic component, presenilin, which lead to increased amyloid ${\beta}$-peptide production, are responsible for early-onset familial Alzheimer's disease. ${\beta}$-amyloid protein precursor-like (APPL) is the Drosophila ortholog of human APP. Here, we created Notch- and APPL-based Drosophila reporter systems for in vivo monitoring of ${\gamma}$-secretase activity. Ectopic expression of the Notch- and APPL-based chimeric reporters in wings results in vein truncation phenotypes. Reporter-mediated vein truncation phenotypes are enhanced by the Notch gain-of-function allele and suppressed by RNAi-mediated knockdown of presenilin. Furthermore, we find that apoptosis partly contributes to the vein truncation phenotypes of the APPL-based reporter, but not to the vein truncation phenotypes of the Notch-based reporter. Taken together, these results suggest that both in vivo reporter systems provide a powerful genetic tool to identify genes that modulate ${\gamma}$-secretase activity and/or APPL metabolism.

Transformation of Bacillus subtilis Protoplast by Recombinant Plasmid DNA (재조합 Plasmid DNA에 의한 Bacillus subtilis의 형질전환)

  • Kim, Sang-Dal;John Spizizen
    • Microbiology and Biotechnology Letters
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    • v.13 no.4
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    • pp.345-348
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    • 1985
  • Recombinant chimeric plasmid constructed with Xba I digested pUBl10 and -pE194 was transformed by polyethylene glycol induced protoplast transformation system into Bacillus subtilis BR 151 on the mannitol regeneration media, and two genes of antibiotics resistance were expressed simultaneously in the transfromant. Transformation frequency of the recombinant plasmid was 6.5 $\times$ 10$^{-5}$ on the mannitol regeneration agar plate containing neomycin and erythromycin. The replication of recombinant plasmid in the recipient cells was confirmed by the alkaline extraction method and agarose gel electrophoresis.

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Expression of Recombinant Rotavirus Proteins Harboring Antigenic Epitopes of the Hepatitis A Virus Polyprotein in Insect Cells

  • Than, Van Thai;Baek, In-Hyuk;Lee, Hee-Young;Kim, Jong-Bum;Shon, Dong-Hwa;Chung, In-Sik;Kim, Won-Yong
    • Biomolecules & Therapeutics
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    • v.20 no.3
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    • pp.320-325
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    • 2012
  • Rotavirus and hepatitis A virus (HAV) spread by the fecal-oral route and infections are important in public health, especially in developing countries. Here, two antigenic epitopes of the HAV polyprotein, domain 2 (D2) and domain 3 (D3), were recombined with rotavirus VP7, generating D2/VP7 and D3/VP7, cloned in a baculovirus expression system, and expressed in Spodoptera frugiperda 9 (Sf9) insect cells. All were highly expressed, with peak expression 2 days post-infection. Western blotting and ELISA revealed that two chimeric proteins were antigenic, but only D2/VP7 was immunogenic and elicited neutralizing antibody responses against rotavirus and HAV by neutralization assay, implicating D2/VP7 as a multivalent subunit-vaccine Candidate for preventing both rotavirus and HAV infections.

Contribution of Perforator Flaps in the Flap Selection for Head and Neck Reconstruction (두경부 재건을 위한 피판 선택에서 천공지피판의 역할)

  • Kim, Jeong Tae
    • Korean Journal of Head & Neck Oncology
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    • v.35 no.1
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    • pp.1-11
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    • 2019
  • In head and neck reconstruction, microsurgeons are faced with various demands, such as thin resurfacing, or three-dimensional reconstruction, and therefore, conventional flap as well as perforator flaps are all useful for the functional and aesthetic reconstruction successfully. Among perforator flaps, the pros and cons of anterolateral thigh perforator flap(ALTp) and thoracodorsal perforator flap(TAp) is compared and selected depending on the surgeon's preference, recipient site condition or patient status. Both flaps can be elevated in a chimeric pattern by combining different aspects of tissue components. Customized or tailored reconstruction as well as 3 dimensional reconstructions are all available with perforator flaps and it is useful in head and neck reconstruction. The most outstanding update in head and neck reconstruction is the perforator based island flap (PBIF) compared to conventional flaps. The conventional local flap has evolved on behalf of the perforator concept and its design becomes more flexible and freestyle with less limitations. Actually, random pattern flap is now a misnomer and most conventional local flaps turn into PBIFs. Finally we can say all conventional donor site becomes universal, depending on the surgeons' preference or idea. Moreover, there is no more "flap of choice" and postoperative results are quite variable by surgeons' ability. Operative procedures and plans are very flexible to freestyle flap. With all these advantages, surgeons should be armed with both conventional and perforator concepts for solving any defects or problems.

The Use of Arteriovenous Bundle Interposition Grafts in Microsurgical Reconstruction: A Systematic Review of the Literature

  • Kareh, Aurora M.;Tadisina, Kashyap Komarraju;Chun, Magnus;Kaswan, Sumesh;Xu, Kyle Y.
    • Archives of Plastic Surgery
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    • v.49 no.4
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    • pp.543-548
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    • 2022
  • Microvascular reconstruction frequently requires anastomosis outside of the zone of injury for successful reconstruction. Multiple options exist for pedicle lengthening including vein grafts, arteriovenous loops, and arteriovenous bundle interposition grafts. The authors performed a systematic review of arteriovenous bundle interposition grafts to elucidate indications and outcomes of arteriovenous grafts in microvascular reconstruction. A systematic review of the literature was performed using targeted keywords. Data extraction was performed by two independent authors, and descriptive statistics were used to analyze pooled data. Forty-four patients underwent pedicle lengthening with an arteriovenous graft from the descending branch of the lateral circumflex femoral artery. Most common indications for flap reconstruction were malignancy (n = 12), trauma (n = 7), and diabetic ulceration (n = 4). The most commonly used free flap was the anterolateral thigh flap (n = 18). There were five complications, with one resulting in flap loss. Arteriovenous bundle interposition grafts are a viable option for pedicle lengthening when free flap distant anastomosis is required. The descending branch of the lateral circumflex femoral artery may be used for a variety of defects and can be used in conjunction with fasciocutaneous, osteocutaneous, muscle, and chimeric free flaps.

Immune cell-derived small extracellular vesicles in cancer treatment

  • Choi, Sung-Jin;Cho, Hanchae;Yea, Kyungmoo;Baek, Moon-Chang
    • BMB Reports
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    • v.55 no.1
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    • pp.48-56
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    • 2022
  • Small extracellular vesicles (sEVs) secreted by most cells carry bioactive macromolecules including proteins, lipids, and nucleic acids for intercellular communication. Given that some immune cell-derived sEVs exhibit anti-cancer properties, these sEVs have received scientific attention for the development of novel anti-cancer immunotherapeutic agents. In this paper, we reviewed the latest advances concerning the biological roles of immune cell-derived sEVs for cancer therapy. sEVs derived from immune cells including dendritic cells (DCs), T cells, natural-killer (NK) cells, and macrophages are good candidates for sEV-based cancer therapy. Besides their role of cancer vaccines, DC-shed sEVs activated cytotoxic lymphocytes and killed tumor cells. sEVs isolated from NK cells and chimeric antigen receptor (CAR) T cells exhibited cytotoxicity against cancer cells. sEVs derived from CD8+ T and CD4+ T cells inhibited cancer-associated cells in tumor microenvironment (TME) and activated B cells, respectively. M1-macrophage-derived sEVs induced M2 to M1 repolarization and also created a pro-inflammatory environment. Hence, these sEVs, via mono or combination therapy, could be considered in the treatment of cancer patients in the future. In addition, sEVs derived from cytokine-stimulated immune cells or sEV engineering could improve their anti-tumor potency.