• Journal of Plant Biotechnology
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• 제38권1호
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• pp.42-48
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• 2011

고등식물의 엽록체 유전공학은 핵 형질전환과 비교해 볼때 여러 가지 독특한 장점을 가진다. 높은 transgene 발현율, 상동 재조합에 의한 site-specific transgene의 삽입으로 인해 유전자의 position effect가 없으며, 단일 형질전환으로 동시에 여러 유전자의 도입이 가능하고 모계 유전으로 인해 화분 방출 위험을 감소시킬 수 있다. 담배 specific한 pCtVG (trnI-Prrn-aadA-mgfp-TpsbA-trnA) 벡터를 이용하여 안정적인 감자 엽록체 형질전환 시스템을 개발하였다. 감자 엽록체 게놈으로 외래유전자의 삽입과 homoplasmic level은 PCR과 Southern blot 분석으로 확인하였다. Northern과 immunoblot 분석 및 GFP fluorescence imaging을 통하여 엽록체 형질전환체의 잎에서 GFP 유전자가 강하게 발현, 축적됨을 알 수 있었다. 본 연구에서 확립된 감자 엽록체 형질전환 시스템을 이용하여 유용 유전자를 도입함으로써 농업적 형질을 개선하거나 고부가가치 단백질을 대량 생산하는 감자를 보다 효율적으로 개발할 수 있을 것이다.

• 한국식물생명공학회:학술대회논문집
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• 한국식물생명공학회 2005년도 추계학술대회 및 한일 식물생명공학 심포지엄
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• pp.156-156
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• 2005

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
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• pp.98-98
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• 2003

Direct gene transfer to mammalian tissues has significant potential for gene therapy and transgenesis. Liposome-mediated in vivo transfection has begun to gain attention as an alternative to viral vectors, and may also be a good mode of transfection in gene transfer. Interestingly, polymerized cationic liposomes are reported to be very stable in the bloods and efficient for in vivo gene transfer. To examine a possible gene delivery in vivo, we investigated the efficacy and safety of the liposome-mediated gene transfer using vein injection in chick or mouse as model animals. The number of injected pGFP-LacZ using either a commercial or home-made liposomes was 8 and 19 at 16 and 7 day of hatch, respectively. One of injected chick of each experiments was analyzed and the rest is being bred. In mouse, 4/22 showed expression of pGFP-LacZ but 8/22 showed no expression and the remaining animals are also being bred. After injection of liposome/pGFP-LacZ complex into wing vein of 7 or 16 day-old chick, pGFP-LacZ was detected in various tissues isolated from not only young chick but also old chick were turned out to possess. exogenous DNA. Transcripts and proteins of the transgene were also detected by RT-PCR or histochemical analysis, respectively. These results suggest that injected DNA were inserted to genome and produced mRNA and proteins in various tissues and may give an important tools for effective gene delivery in gene therapy or transgenesis.

• Journal of Plant Biotechnology
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• 제33권3호
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• pp.185-194
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• 2006

엽록체는 숙주세포에 잡아먹힌 (식균작용) 남세균이 숙주세포와 공생관계를 형성하여 온 것으로 간주된다. 엽록체 게놈은 정적이라고 이해하고 있지만 형질전환을 통하여 상동염기가 도입되면 이와는 반대로 intramolecular homologous recombination에 의해 subgenomic circle을 만드는 등 매우 다이나믹하다는 것이 최근에 증명되고 있다. 고등식물의 엽록체 형질전환은 핵 형질전환에서 기대할 수 없는 여러 이점을 제공한다. 예컨대, transgene의 발현율을 높일 수 있고, transgene들을 polycistronic하게 발현할 수 있으며, 도입된 transgene이 모계유전을 하게 된다는 것 등이다. 담배는 엽록체 형질전환의 모델 식물로 사용되어 왔으나 최근에는 벼, 대두, 면화 등 다른 주요 작물의 형질전환도 가능하게 되었다. 엽록체 형질전환된 작물은 미생물을 이용하여 고부가가치 단백질을 생산하는 생물반응기를 향후 대체할 수 있게 될 것이다.

• 대한의생명과학회지
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• 제17권3호
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• pp.185-190
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• 2011

Adenovirus type 5 (Ad5) vectors have been used for gene transfer to a wide variety of cell types in vivo and in vitro. The advantages of adenovirus vectors include the high titer of virus readily obtained in large scale preparations, their ability to transduce dividing and non dividing cells, and the high level of transgene expression. Since adenovirus vectors do not integrate in host cell DNA, there is a lack of insertional mutagenesis. However, many human tumor cells lack expression of the adenovirus 5 receptors and contain areas of hypoxia. In order to identify the pattern of replication and gene expression of oncolytic adenovirus in hypoxic condition, multiple different fiber modified Ads (Ad5F/S11, Ad5F/S35, Ad5F/K7, Ad5F/K21, and Ad5F/RGD) was compared. The replication of all fiber modified adenovirus was inhibited in hypoxic condition in HEK 293 cells, but gene expression has variety on different tumor cell lines and the level of coxackievirus and adenovirus receptor (CAR) expression. These data suggest that CAR expression pattern and hypoxic condition of tumor are considered for optimal oncolytic adenovirus application.

• 한국식물생명공학회:학술대회논문집
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• 한국식물생명공학회 2005년도 추계학술대회 및 한일 식물생명공학 심포지엄
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• pp.390-390
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• 2005

• Journal of Plant Biotechnology
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• 제6권1호
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• pp.15-19
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• 2004

The expression of a chimeric transgene (nos-npt II) has been examined in 9 years old transgenic poplars (Populus koreana x P. nigra) growing in a nursery. The expression of the gene in twenty six independentely transformed plants were examined by 1) enzyme (NPT II) assay, 2) RT-PCR, and 3) resistance to kanamycin. High NPT II activities in young leaves of all the transformed plants were found even without a selection pressure for antibiotics for 9 years. However, the activity varied with the positions of leaves in the stem in that young leaves showed higher activity than did mature tissues. When leaf segments were cultured in the presence of 150 mg/l kanamycin, only those from young leaves produced vigorously growing callus. However, as in the case of NPTII assay, the leaf segments from mature leaves did not form callus well on the media. RT-PCR with nptII specific primers also showed that amplification products were observed only when RNAs from young tissues were used. The total RNA gel showed that while RNA in young leaves are relatively stable and in a large quantity, those in old leaves were mostly degraded. All the above results suggest that the gene is transcriptionally active only in young tissue even though it is attached to a constituitive promoter. Therefore, the expression of foreign gene in poplar plants seemed to be affected by the metabolic state of the cells and thus vary greatly with the developmental stages and the age of tissue.

• The Plant Pathology Journal
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• 제27권4호
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• pp.310-314
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• 2011

Nonstructural protein 3 (NS3) encoded by RNA3 of Rice stripe virus (RSV), known to be a suppressor of gene silencing, was cloned and sequenced. The cloned NS3 gene is composed of 636 nucleotides encoding 211 deduced amino acids, and showed a high degree of similarity with the equivalent genes isolated from Korea, Japan and China. The NS3 gene promoted the enhancement of transient gene expression and suppressed transgene co-silencing. In the transient GFP expression via agroinfiltration, GFP expression was dramatically enhanced in terms of both protein yield and expression period in the presence of NS3. The highest accumulation of GFP protein reached to 6.8% of total soluble proteins, which corresponded to a two-fold increase compared to that obtained in the absence of NS3. In addition, NS3 significantly suppressed the initiation of GFP co-silencing induced by the additive GFP infiltration in GFP-transgenic Nicotiana benthamiana. The NS3 gene was also found to be a stronger suppressor than Cucumber mosaic virus 2b. These observations are believed to be derived from the strong suppressive effect of NS3 on gene silencing, and indicate that NS3 could be used as an effective enhancer for the rapid production of foreign proteins in plants.

• 한국수정란이식학회지
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• 제30권3호
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• pp.225-228
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• 2015

Understanding the behavior of transgenes introduced into oocyte or embryos is essential for evaluating the methodologies for transgenic animal production. To date, many studies have reported the production of transgenic pig embryos with, however, low efficiency in environment of blastocyst production. The aim of present study was to determine the expression and duration of transgene transferred by intracytoplasmic sperm injection-mediated gene transfer (ICSI-MGT). Embryos obtained from the ICSI-MGT procedure were analysed for the expression of GFP and then for the transmission of the transgene. Briefly, fresh spermatozoa were bound to exogenous DNA after treatment by Triton X-100 and Lipofectin. When ICSI-MGT was performed using sperm heads with tails removed, the yield of blastocyst (25.3%), treated with Lipofectin (18.8%) and Triton X-100 (19.2%) were observed. Treatments of Lipofectin or Triton X-100 did not further improve the rates of blastocysts. Moreover, the apoptosis rates of embryos were obtained from the control and LIpofectin groups (8.7%, 9.7%, respectively), but were significantly higher in the Triton X-100 group (13.0%). Our results demonstrated that ICSI-MGT caused minimal damage to oocytes that could develop to full term. Moreover, the embryos derived by ICSI-MGT have shown prolonged exogenous DNA expression during preimplantation stage in vivo. However, more efforts will be required to improve the procedures of both sperm treatments cause of high frequency of mosaicisms.

• 한국어업기술학회:학술대회논문집
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• 한국어업기술학회 2002년도 추계 수산관련학회 공동학술대회발표요지집
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• pp.183-184
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• 2002

The use of transgenic fish has so far been chiefly limited by the lack of predictable, strong, tissue specific, and position-independent expression of transgenes. For genetic analysis, expression of a marker transgene, easily screenable in the living fish, could facilitate studies of gene targeting, insertional mutagenesis, lineage, and mutational analysis. (omitted)