Delta 15 desaturase 유전자 억제에 의해 알파리놀렌산 함량이 낮은 들깨 육성

김경환(Kyung-Hwan Kim),이경렬(Kyeong-Ryeol Lee),김정봉(Jung-Bong Kim),이명희(Myoung Hee Lee),이은경(Eungyeong Lee),김년희(Nyunhee Kim),이홍석(Hongseok Lee),김송림(Song Lim Kim),백정호(JeongHo Baek),최인찬(Inchan Choi),지현소(Hyeonso 한국육종학회 2018 한국육종학회지 Vol.50 No.4

Perilla is an oilseed crop cultivated in Korea since ancient times. Due to the high α-linolenic acid content in perilla, perilla seed oil can easily become rancid. α-Linolenic acid is synthesized by two enzymes, endoplasmic reticulum-localized Δ15 desaturase (FAD3) and chloroplast-localized Δ15 desaturase (FAD7) in vivo. In order to lower the α-linolenic acid content of the seed oil without disturbing plant growth, we tried to suppress the expression of only the FAD3 gene using RNA interference, whilst maintaining the expression of the FAD7 gene. Seventeen transgenic plants with herbicide (Basta TM ) resistance were obtained by Agrobacterium-mediated transformation using hypocotyls of perilla plants. The transgenic plants were firstly confirmed by treatment with 0.3% (v/v) Basta TM herbicide, and the expression of FAD3 was measured by Northern blot analysis. The α-linolenic acid content was 10-20%, 30-40%, and 60% in two, seven, and three of the twelve T1 transgenic perilla plants which had enough seeds to be analyzed for fatty acid composition, respectively. Analysis of the fatty acid composition of T2 progeny seeds from T1 plants with the lowest α-linolenic acid content showed that the homozygous lines had 6-10% α-linolenic acid content and the heterozygous lines had 20-26% α-linolenic acid content. It is expected that the reduction in α-linolenic acid content in perilla seed oil will prevent rancidity and can be utilized for the production of high-value functional ingredients such as high γ-linolenic acid.

식물의 산업용 지방산 생산을 위한 오일합성 유전자의 기능과 이용 전망

김현욱,이경렬,박종석,노경희,김순희,김종범,Kim, Hyun-Uk,Lee, Kyeong-Ryeol,Park, Jong-Sug,Roh, Kyung-Hee,Kim, Sun-Hee,Kim, Jong-Bum 한국식물생명공학회 2010 식물생명공학회지 Vol.37 No.2

식물 종자오일의 지방산은 인간에 필수 지방산을 공급하는 식용 및 생필품 생산에 필요한 다양한 산업원료로 사용된다. 식물 오일의 식용 및 산업용 적합성과 경제성을 극대화하기 위해 유전공학에 의한 종자오일의 양과 지방산 조성 변형을 위한 대사조절연구가 계속 진행되고 있다. 하지만 식물에 일반적으로 존재하지 않는, 산업적으로 유용한 특이지방산의 합성과 종자오일로의 축적은 한계가 있음이 알려져 있다. 그 이유는 재배가 용이하며 생산성이 높은 오일식물의 acyltransferase가 특이지방산에 대한 기질특이성이 떨어지며 또한 특이지방산에 대한 세포막지질에서 종자오일로 전환시키는 편집기작 (editing mechanism)이 없기 때문으로 사료된다. 최근에 모델식물의 종자오일의 축적에 관여하는 acyltransferase에 관한 유전자들이 클로닝되었고, 특이지방산이 합성되는 인지질에서의 편집기작에 관여하는 acyltransferase 유전자들이 밝혀져 이들 유전자들의 정보를 이용하여 특이지방산을 효과적으로 생산.증진할 수 있는 기술이 개발될 수 있을 것으로 기대한다. 피마자오일의 주성분인 산업용 특이지방산인 리시놀레인 지방산을 오일식물에서 생산하기 위해 이에 관여할 것으로 추정되는 11개의 acyltransferase 유전자를 피마자 유전체 데이터베이스에서 존재함을 확인하였다. 이들 유전자들의 도입에 의해 형질전환 식물이 갖고 있지 않은 리시놀레인산에 대한 기질 특이성을 부여하여 종자오일 내의 특이지방산의 생산을 증가시킬 것으로 기대된다. Fatty acids in seed oil from plants are essential for human nutrients and have been used for industrial purpose. The growing demands of seed oil as food resources and feedstocks for industrial uses have attempted to modify fatty acid composition and to increase oil content in transgenic plants. However, production of unusual fatty acids in transgenic plants are limited, which is not synthesized the level same as original plants. This bottleneck was common for production of several unusual fatty acids in transgenic plants and suggests that there is different for substrate preference in oil metabolic pathway enzymes between host oil plants and original wild plants. Review of acyltransferases involved in acyl-editing and seed oil accumulation of oil plant and wild-plant producing unusual fatty acids will design strategies to maximize the production of unusual fatty acids in transgenic plants. In here, we identified eleven acyltransferase genes in castor based on sequence homology, which will be useful to increase hydroxy unusual fatty acids in transgenic plants.

RNA 결합 단백질과 유전자 발현조절

노경희 ( Kyung Hee Roh ),강한철 ( Han Chul Kang ),김종범 ( Jong Bum Kim ),김현욱 ( Hyun Uk Kim ),이경렬 ( Kyung Ryeol Lee ),김순희 ( Sun Hee Kim ) 한국응용생명화학회 2015 Journal of Applied Biological Chemistry (J. Appl. Vol.58 No.3

RNA 결합 단백질들이 유전자 조절의 다양한 범위에 작용한다는 사실이 아주 중요하다. 유전자의 전사에 관련된 유전자 조절이 많이 연구가 되었어도 RNA 의 조절에 관한 연구는 상대적으로 부진한 편이다. RNA 결합 단백질들은 RNA 와 관련되는 각종 과정, 예를 들면 전사, pre-mRNA splicing, polyadenylation, 수송,위치화, 번역, 분해 및 구조의 유지 등 다양한 범위에서 작용을 하고 있다. RNA 결합 단백질들의 많은 부분들이 아직 잘 알려지지 않고 있으며 유전자 발현에 대해 더 잘 이해하기 위해 이러한 부분의 연구가 더 수행되어야 한다. 최근에 유전학, 생화학, 및 유전자들의 생물정보학의 발달 등으로 인하여, RNA 결합 단백질들의 다양한 분야들이 알려지고 있으며 이러한 부분들이 많은 관심을 받고 있다. The role of RNA-binding proteins (RBPs) to regulate expression of genes seems to be very important. RBPs play important roles in RNA related bioprocess such as transcription, pre-mRNA splicing, polyadenylation, transport, localization, translation, turn over and maintenance of structure. Despite of many researches on RNA binding proteins, detailed mechanisms of these proteins have not been fully understood. It seems that many parts of RBPs remains unknown and should be characterized for the better understanding of gene expression. Recently, genetic, biochemical, and bioinformatic analysis of genomes revealed a vast array of RBPs and many parts are interesting to understand bioprocessing including gene expression.

영산 유채를 이용한 형질전환체 생산

노경희 ( Kyung Hee Roh ),곽보경 ( Bo Kyoung Kwak ),김현욱 ( Hyun Uk Kim ),이경렬 ( Kyeong Ryeol Lee ),김순희 ( Sun Hee Kim ),서미정 ( Mi Chung Suh ),김효진 ( Hyo Jin Kim ),김종범 ( Jong Bum Kim ) 한국응용생명화학회 2011 Journal of Applied Biological Chemistry (J. Appl. Vol.54 No.1

To improve genetic transformation of Brassica napus winter cultivar ``Youngsan``, factors influencing shoot regeneration and transformation from cotyledonary petioles were investigated. Shoot induction was enhanced in the combination of 0.5 mg/L NAA and 2~4 mg/L kinetin. Silver nitrate was essential for successful shoot regeneration, ranging from 5 to 9 mg/L. The addition of GA3 promoted plant regeneration. Among the tested Agrobacterium strains, co-cultivation times, and antibiotic selection regimes, choice of appropriate Agrobacterium strain was the most critical factor for efficient transformation of B. napus cv. ``Youngsan``. The EHA105 succinamopine strain was the most efficient and the maximum transformation efficiency was 26.8%. Transgenic shoots were selected on 10 mg/L phosphinothricin (PPT) containing media. The transgenic plants expressing bar and gus genes were resistant for commercial herbicide Basta and stained with X-Gluc. Southern blot hybridization indicated that the presence of one to three gus gene copies per genome and inheritance of the gus gene into the T1 generation.

Microarray 분석을 이용한 유채 종자성숙단계별 유전자 발현 양상

노경희 ( Kyung Hee Roh ),박종석 ( Jong Sug Park ),김종범 ( Jong Bum Kim ),김현욱 ( Hyun Uk Kim ),이경렬 ( Kyeong Ryeol Lee ),김순희 ( Sun Hee Kim ) 한국응용생명화학회(구 한국농화학회) 2012 Journal of Applied Biological Chemistry (J. Appl. Vol.55 No.4

We observed that oil began to accumulate at 25 seed days after flowering (DAF) and reached the maximum potential at 35 seed DAF of oilseed rape, and the greatest weight of 100 seeds was obtained at 35 seed DAF. To survey a broad analysis of gene expression in developing embryos of Brassica napus, the Bn 300k microarray have been constructed. The Bn 300k Microarrary was designed from 80,696 unigenes clustered from 543,448 ESTs and 780 cDNA at NCBI. These arrays have been hybridized in a series of experiments with probes derived from seeds and leaf of B. napus. Approximately 8.5% of the 7,000 genes were expressed as ratios 2-fold higher in seed (25 DAF) than leaves and 0.4% at ratios 10. Also we observed that storage and cell differentiationrelated genes were highly expressed at 10 DAF, whereas energyrelated genes including fatty acid metabolism were increased up depending on seed maturation using Microarray, which was confirmed by reverse transcriptase polymerase chain reaction. These results suggest that B. napus arrays provide a very useful data set of seed-specific expression that can be further analyzed by examination of the promoter regions of these genes and help our understanding of the complex regulatory network in developing seeds.

의료용 단백질 생산을 위한 바이러스 운반체 시스템

김순희 ( Sun Hee Kim ),강한철 ( Han Chul Kang ),노경희 ( Kyung Hee Roh ),김현욱 ( Hyun Uk Kim ),이경렬 ( Kyeong Ryeol Lee ),김원용 ( Won Yong Kim ),박종화 ( Jong Hwa Park ),정인식 ( In Sig Chung ),김종범 ( Jong Bum Kim ) 한국국제농업개발학회 2015 韓國國際農業開發學會誌 Vol.27 No.2

Most of pharmaceutical proteins are usually produced by the animal, insect cell and yeast culture. The proteins must be produced in strictly controlled-facilities, which is one of major causes for increasing production cost. So, in spite of increasing demand on the proteins in worldwide, their generalization has been very restrictive. To decrease the production cost, many scientists have been interested in production of pharmaceutical proteins by using the plant system for a long time. The plant system must be alternative production means, for it is easily scalable, more production cost effective, and safer than animal system. For this purpose, several techniques have been developed: stable expression by transformation of nuclear and chloroplast genome and transient expression by viral vectors. Among them, the deconstructed viral vector system has made it possible to produce recombinant proteins massively, rapidly, and transiently. So, the plant viral expression systems might be a suitable platform for production of pharmaceutical proteins in plants. In this review, we will describe the plant viral vector systems and their application.

히스톤의 변이와 이와 관련된 기능적 측면

강한철 ( Han Chul Kang ),김종범 ( Jong Bum Kim ),노경희 ( Kyung Hee Roh ),김현욱 ( Hyun Uk Kim ),이경렬 ( Kyung Ryeol Lee ),김순희 ( Sun Hee Kim ) 한국응용생명화학회(구 한국농화학회) 2014 Journal of Applied Biological Chemistry (J. Appl. Vol.57 No.4

크로마틴은 DNA 구조를 다시 결정해주는 것과 같은 존재로 각종 신호에 폭넓게 반응한다. 크로마틴의 중요한 변화는 이러한 조절을 위한 히스톤의 변이이다. 이러한 변화들에 대한 지식이 점점 축적되고 있으며 이러한 반응의 복잡성이 점점 더 명확히이해되고 있다. 히스톤의 변화가 대부분의 생명체의 반응에 있어서 DNA 의 발현 또는 억제를 통하여 중요한 역할을 한다는 사실이 명확해지고 있다. Nucleosome 의 표면은 각종 변화를 수용할 수 있다. 크로마틴 변화는 크로마틴 수축을 제거하거나 또는 비히스톤 단백질들을 불러 모으는 과정을 통하여 작용될 수 있다. 히스톤 변이를 매개로 하는 이러한 많은 조절들이 유전적으로 보존되어 전달되는 것으로 추측된다. 따라서 히스톤변이는 동물, 식물 또는 미생물 세계의 기본적인 생물학적 반응과 상당히 밀접한 관계가 있다. 히스톤 변이가 제대로 이루어지지 않을 경우 크로모좀의 응축 또는 이완이 제대로 않되며 결국은 발생, 성숙, 생물체 방어 등 다방면에 대해 기능을 제대로 수행하지 못한다. Chromatin is an instructive DNA structure that canwidely respond to external signals. An important change of chromatin is the modifications of histone for this regulation. There are accumulating lists of these modifications and the complexity of their action is gradually understood. It is evident that histone modifications play important roles in most biological processes that are involved in the expression or repression of DNA. The surface of nucleosomes is susceptible to multiplicity of modifications. Chromatin modifications can play either by eliminating chromatin contacts or by recruiting non-histone proteins to chromatin. Many of these regulations seem to be epigenetically inherited. Thus, histone modifications are closely correlated with many fundamental biological processes in animal, plant and microbial kingdoms. Failures of histone modification lead, in general, to defective chromosome condensation or decondensation, impeding many biological functions including development, maturation, and protection against various diseases.