• Journal of the Korean Society of Food Science and Nutrition
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• v.29 no.6
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• pp.1030-1034
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• 2000

The ovalbumin, a most sensitive egg white protein to irradiation was purified from irradiated hen's eggs. Eggs were irradiated in their shells to 0~7 kGy. To investigate for a practical use in identifying of irradiated eggs, competitive ELISA using ovalbumin was peformed. The binding activity of ovalbumin to anti-ovalbumin IgG was reduced in a dose-dependent manner by irradiating up to 7 kGy, and consider-ably lowered after irradiating at 7 kGy. The concentration of 50% inhibition of ovalbumin to IgG was increased to 1.5~3.7 times in an irradiation dose-dependent relationship. SDS-PAGE of ovalbumin showed that the partial breakdown of ovalbumin was induced by irradiation. The lowering of binding activity was probably due to the partial breakdown of ovalbumin by irradiation. These results demonstrated that the ELISA should be quite useful and effective methods for the identification of irradiated eggs.

• KOREAN POULTRY JOURNAL
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• v.46 no.10
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• pp.132-133
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• 2014

생식선 줄기세포 원천기술을 이용한 형질전환 조류 생산 시스템과 고효율의 유전자적 중 기술인 유전자가위법(TALEN)을 도입함으로써 기초연구 및 단기간내 새로운 가금품종 개발이 가능하게 되었다. 또한, 신약개발 및 치료물질 대량생산을 위한 형질전환가금품종 개발에 응용될 수도 있어 축산과 더불어 의약, 약학 등 매우 다양한 분야에서 가금의 활용 범위를 넓힐 수 있을것으로 기대된다. 개발된 기술은 계란성분 조절을 통한 기능성 식품 및 단백질-신약을 포함한 신물질 생산을 목적으로 하는 지식기반 생명산업의 획기적인 발전을 유도할 수 있다. 이에 유전자 가위 기법을 활용한 오브알부민 생산 유전자를 제거한 유전자 변형 닭 생산에 관한 주요내용을 소개코자 한다.

• Proceedings of the Korea Society of Poultry Science Conference
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• 2006.11a
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• pp.80-81
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• 2006

닭의 유전자 지도가 밝혀지고 그와 관련한 생물학적 연구들이 활발히 이루어지면서 닭을 생체 반응기나 질병 모델 동물로 이용하기 위한 연구가 많이 진행되고 있다. 이 중 닭을 생체 반응기로 이용하기 위해서는 많은 양의 단백질을 생산하는 난관에 대한 연구가 필수적이다. In vivo와 in vitro에서 난관 특이적 프로모터에 의한 외래 유전자의 발현에 대한 연구를 하였고 유전자를 전이하는 방법으로는 렌티 바이러스 시스템을 이용하였으며, 프로모터는 난관 특이적 프로모터인 오브알부민 프로모터 (5‘ 조절 부분의 1.4kb)와 RSV 프로모터를 이용하였다. 리포터 유전자로는 형광발현 단백질 (enhanced green fluorescence protein, EGFP)을 이용해서 마우스 배아 섬유아세포, 닭 배아 섬유아세포, 난관 상피 세포에서 발현을 유도해서 조직 특이적 발현 여부를 확인하였다. 그 결과 RSV 프로모터는 모든 세포에서 발현하였으나, 오브알부민 프로모터에 의한 리포터 유전자의 발현은 난관 상피 세포에서는 특이적으로 발현하였다. 이와 같은 연구는 산란계를 이용해서 난관으로부터 효율적인 생리 활성 물질을 생산하기 위한 가능성을 보여주었다.

• Korean Journal of Poultry Science
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• v.46 no.1
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• pp.17-24
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• 2019

Chickens have been considered as well-defined animal bioreactor. The optimized ovalbumin promoter is essential for recombinant protein production in transgenic chicken. Here we try to compare the activity and identify the effect of estrogen on ovalbumin promoter according to each promoter length with estrogen response element (ERE) existence. We cloned two (2.8 and 5.5 kb) ovalbumin promoters that the 5.5 kb contained the ERE but the 2.8 kb did not, and these two promoters were cloned to pGL4.11 vector. Additionally, we constructed another pGL4.11 vector containing of the 4.4 kb (with ERE) ovalbumin promoter deleted with 1 kb between ERE region and the 2.8 kb promoter. For reporter assay, HeLa, MES-SA, LMH/2A, and cEF cells were transfected with all the pGL4.11 vectors. The comparative analysis showed that the mutated 4.4 kb promoter has more potent activity than the 2.8 and 5.5 kb promoters in HeLa, MES-SA, and LMH/2A cells. However, there is no significant difference in cEFs. Also, these cells transfected with the mutated 4.4 kb promoter were treated with the $17{\beta}$-estradiol (0~3,000 nM) and HeLa, MES-SA, and LMH/2A cells showed estrogen responsibilities, but cEFs did not. Besides, the mutated 4.4 kb promoter has still higher activity than the 2.8 and 5.5 kb promoter, and there is no transcriptional induction effect in 2.8 kb promoter at 500 nM estrogen that is blood concentration of laying hens. Hence our study strongly suggested that the mutated 4.4 kb promoter is considered as one of the most efficient length for generating transgenic chicken.

• Journal of Life Science
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• v.27 no.4
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• pp.450-455
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• 2017

Codonopsis lanceolata (Campanulaceae) has been widely used in traditional medicine and is considered to have medicinal properties to treat diseases and symptoms such as bronchitis, coughs, spasm, edema, hepatitis, colitis, and lung injury. In order to investigate whether native Codonopsis lanceolata extract alleviates ashmatic symptoms in vivo, we first carried out various antioxidant activities by 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging, ferric reducing antioxidant power (FRAP), and cupric reducing antioxidant capacity (CUPRAC) assays. The antioxidant activities were increased by adding Codonopsis lanceolata extract in a concentration-dependent manner which compared to ascorbic acid as a positive control. Histological studies using an ovalbumin-induced animal model exhibited potent anti-inflammatory potential by decreasing immuno-responsive cells in the lung by the extract by confirming H&E and PAS staining. It is revelaed that further immunihistochemical analysis showed anti-ashmatic capabilities by assessing histamine, IL-31, and MMP-9 expressions. The level of IL-13 expression in Codonopsis lanceolata extract-treated group was decreased upto 73.7% compared to control, whereas that of total cells and eosinophil counting in Codonopsis lanceolata extract-treated group was diminished to 73.5% and 80.9%, respectively. These results collectively indicate that the C. lanceolata extract ameliorates asthmatic symptoms effectively in an ovalbumin-challenged mice model, in that the extract can be used for the development of an anti-asthmatic food ingredient.

• Applied Biological Chemistry
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• v.51 no.4
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• pp.276-280
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• 2008

To elucidate the effects of ultraviolet (UV) irradiation on molecular properties of ovalbumin, the molecular weight profile, secondary structure and tertiary structure of proteins were examined after irradiation by UV with 254 nm wavelength for 4, 8, 16 and 32 hrs, respectively. UV irradiation of protein solution caused the disruption on the native state of protein molecules. SDS-PAGE and gel permeation chromatography indicated that radiation caused initial fragmentation of polypeptide chains and as a result subsequent aggregation due to cross-linking of protein molecules. Circular dichroism (CD) study showed that UV irradiation caused the change on the secondary structure resulting in decrease of helical structure or compact denature on structure of protein depending on irradiation period. Fluorescence spectroscopy indicated that irradiation quenched the emission intensity excited at 280 nm. These results suggest that UV irradiation affect the molecular properties of ovalbumin and may have potential as a means to change the antigenicity of protein allergen.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.13 no.12
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• pp.6208-6214
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• 2012

The most essential but missing components to understand and use toxic substances from marine microalgae are developing the fast, easy and economical determining technology for detecting it. In this paper we produced the antibodies against saxitoxin (STX). Mariculture keyhole limpet hemocyanin (mcKLH) and ovalbumin (OVA) were used as carrier proteins. mcKLH-STX conjugates were injected into the peritonial cavity of BALB/c mouse for immunization. After bleeding from mouse, anti-STX antiserum was isolated. Indirect enzyme-linked immunosorbent assays (ELISA) was performed to determine antiserum titer using the microtiter plate coated with free STX and OVA-STX. A goat anti-mouse IgG-phosphatase conjugate was used as secondary antibody to enable chromogenic reaction. Reactions of anti-STX antiserum were very specific on the OVA-STX and free STX. Sensitivity of anti-STX antiserum on STX was very high and STX detection limit was to be 64.9 ng/kg for indirect ELISA.