• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2004년도 춘계학술발표대회
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• pp.194-194
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• 2004

Tissue-specific expression of the desired gene product in the targrt tissue is central to the concept of bioreactor. One approach is to use a tissue-specific promoter to drive desired gene. To investigate the feasibility of tissue-specific gene expression for bladder using the porcine uroplakin(UPII) promoter and its transcriptional control the efficacy of this promoter as well as well as fragments in regulating gene expression were cell lines using DNA transfection. (omitted)

• 식물조직배양학회지
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• 제25권4호
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• pp.237-243
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• 1998

야생종 토마토의 자가불화합성에 관여하는 S RNase유전자의 조직특이적 발현 양상을 조사하기 위하여 $\textrm{S}_{11}$ 및 $\textrm{S}_{12}$ allele에 속하는 RNase 유전자의 Promoter영역에 대한 염기 서열을 비교 분석한 결과, 전사개시점에서 상류측으로 350-500bp사이에서 양쪽 allele의 Promoter간에 상동성을 나타내는 3부분과 direct repeat sequence를 발견하였다. Promoter영역에서 이러한 부분이 S RNase 유전자가 화주특이적으로 발현하는데 영향을 줄 것으로 예상하고, 이들 영역을 중심으로 6종류의 deletion fragment를 만들어 GUS 유전자에 연결하여, 토마토의 생식조직에 microprojectile bombardment를 수행하였다. 그 결과 토마토 자가불화합성에 관여하는 S RNase 유전자의 promoter는 TATA box를 포함한 127 bP만으로도 화주조직 특이적 발현을 조절할 수 있었다. 또한 S RNase 유전자의 promoter영역내에는 토마토 화변, 자방과 심피조직들에서 negative 혹은 positive로 유전자의 발현을 유도하는 부분이 발견되었다.

• 한국정보과학회:학술대회논문집
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• 한국정보과학회 1999년도 가을 학술발표논문집 Vol.26 No.2 (2)
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• pp.12-14
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• 1999

Promoter는 transcript start site 앞부분에 위치하여 RNA polymerase가 높은 친화성을 보이며 바인당하는 DNA상의 특별한 부위로서 여기서부터 DNA transcription이 시작된다. function이나 tissue-specific gene들의 그룹별로 그 promoter들의 특이한 패턴들의 조합을 발견함으로써 Specific한 transcription을 조절하는 것으로 알려져 있어 promoter로 인한 그 gene의 정보를 어느 정도 알 수가 있다. 사람의 housekeeping gene promoter들을 EPD(eukaryotic promoter database)와 EMBL nucleic acid sequence database로부터 수집하여 이것들 간에 의미 있게 나타나는 모든 패턴들을 optimization algorithm으로 알려진 genetic algorithm을 이용해서 찾아보았다.

• International Journal of Industrial Entomology and Biomaterials
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• 제25권1호
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• pp.111-114
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• 2012

In previous studies we have shown that a sw17255 gene was expressed in hemocyte-specific tissues of the silkworm, Bombyx mori L. (Lepidoptera: Bombycidae). It was verified that the sw17255 core promoter region contains elements that regulate the expression of this gene in hemocyte tissue; the selected promoter region spans nucleotides -1 to -2,112 upstream of the start codon. Each of the luciferase reporter gene expression vectors under the control of 4 different kinds of promoter candidates, (-2,112/-1), (-1,640/-1), (-1,169/-1) and (-579/-1), and the control reporter plasmid DNA, were introduced into B. mori larval coelom by direct injection using a syringe. The promoter candidate [E] (-579/-1) showed more than 1.67 fold transcriptional activity compared to control promoter activity. Higher productivity of an expressed gene in the transgenic silkworm by this promoter combination could be achieved in the near future. The foreign recombinant protein could be easily harvested from the blood of the transgenic silkworm.

• Reproductive and Developmental Biology
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• 제40권1호
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• pp.7-13
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• 2016

Effectiveness of transgene transfer into genome is crucially concerned in mass production of the bio-pharmaceuticals using genetically modified transgenic animals as a bioreactor. Recently, the mammary gland has been considered as a potential bioreactor for the mass production of the bio-pharmaceuticals, which appears to be capable of appropriate post-translational modifications of recombinant proteins. The mammary gland tissue specific vector system may be helpful in solving serious physiological disturbance problems which have been a major obstacle in successful production of transgenic animals. In this study, to minimize physiological disturbance caused by constitutive over-expression of the exogenous gene, we constructed new retrovirus vector system designed for mammary gland-specific expression of the hEPO gene. Using piggyBac vector system, we designed to express hEPO gene under the control of mammary gland tissue specific and lactogenic hormonal inducible goat ${\beta}$-casein or mouse Whey Acidic Protein (mWAP) promoter. Inducible expression of the hEPO gene was confirmed using RT-PCR and ELISA in the mouse mammary gland cells treated with lactogenic hormone. We expect the vector system may optimize production efficiency of transgenic animal and reduce the risk of global expression of transgene.

• Journal of Plant Biotechnology
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• 제42권4호
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• pp.356-363
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• 2015

나무의 유전 공학적 연구를 위해서는 목본 고유의 유전자 및 프로모터 연구가 필수적이다. 우리는 포플러(P. alba ${\times}$ P. glandulosa)의 Pagns-LTP 유전자의 867 bp 프로모터를 분리하였고, ${\beta}$-glucuronidase (GUS) reporter 유전자를 이용한 프로모터의 형질전환 포플러를 제작하여 특성 분석하였다. Pagns-LTP 유전자는 어린뿌리에서 강하게 발현되었고 어린잎에서는 약하게 발현되었으며, 그밖에 다른 조직에서는 발현되지 않았다. 또한, 프로모터의 활성은 뿌리와 어린잎에서 한정되었으며 어린뿌리의 세포 전체에서 강한 활성을 나타내었다. 이에 포플러 ns-LTP 프로모터 내의 cis-element를 조사하고 현사시나무에서 Pagns-LTP 프로모터를 분리한 후 활성을 분석하였다. 프로모터 내의 cis-element를 분석한 결과, 조직 특이적 발현과 호르몬 및 스트레스에 반응하는 다양한 cis-element가 존재함을 확인하였다. 이를 통해 포플러의 ns-LTP는 생장뿐만 아니라, 스트레스에도 관여할 것이라고 추측할 수 있었다. 본 연구는 목본의 유전자 기능 분석 및 다양한 응용 연구를 위해 유용하게 이용될 수 있는 도구로서의 가능성을 제시하였다.

• 미생물학회지
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• 제30권6호
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• pp.488-494
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• 1992

Three clones containing mouse CD4 gene were prepared using AKR genomic cosmid library. The role of 6, 500 bp 5' flanking region of the first exon of the AKR CD4 gene in tissue or developmental stage specific expression of CD4 has been studied. The deletion constructs containing various amounts of CD4 5' flanking sequences were prepared, and they were transfected into the cell lines representing different cell types or developmental stages of CD4 expression. Study of the reporter gene expression revealed that at least 1, 700 bp of 5' flanking region did retain promoter activity for CD4 expression. This area did not seem to contain enhancer activity for a full expression of CD4. However, the putative promoter interacted with other tissue specific enhancer sequence and showed the tissue specificity of the enhancer element.

• Reproductive and Developmental Biology
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• 제36권3호
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• pp.207-212
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• 2012

The shortage of human organs for transplantation has induced the research on the possibility of using animal as porcine. However, pig to human transplantation as known as xeno-transplantation has major problem as immunorejection. Recently, the solutions of pig to human xenotransplantation are commonly mentioned as having a genetically modification which include alpha 1, 3 galatosyl transferase knockout (GTKO) and immune-suppressing gene transgenic model. Unfortunately, the expression level of transgenic gene is very low activity. Therefore, development of gene overexpression system is the most urgent issue. Also, the tissue specific overexpression system is very important. Because most blood vessels are endothelial cells, establishment of the endothelial-specific promoter is attractive candidates for the introduction of suppressing immunorejection. In this study, we focus the ICAM2 promoter which has endothelial-specific regulatory region. To detect the regulatory region of ICAM2 promoter, we cloned 3.7 kb size mini-pig ICAM2 promoter. We conduct serial deletion of 5' flanking region of mini-pig ICAM2 promoter then selected promoter size as 1 kb, 1.5 kb, 2 kb, 2.5 kb, and 3 kb. To analyze promoter activity, luciferase assay system was conducted among these vectors and compare endothelial activity with epithelial cells. The reporter gene assay revealed that ICAM2 promoter has critical activity in endothelial cells (CPAE) and 1 kb size of ICAM2 promoter activity was significantly increased. Taken together, our studies suggest that mini-pig ICMA2 promoter is endothelial cell specific overexpression promoter and among above all size of promoters, 1 kb size promoter is optimal candidate to overcome the vascular immunorejection in pig to human xenotransplantation.

• 생명과학회지
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• 제15권2호
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• pp.176-181
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• 2005

Calmodulin 유전자의 발현 조절을 연구하기 위해, 벼 calmodulin promoter (RCaM-2)를 분리하여 GUS (report 유전자)에 융합하였다. GUS 활성은 정단조직, 근단 및 관다발 영역과 같은 성장조직에서 높게 발현되었다. 줄기와 페티올의 transverse 절단부위 GUS 활성은 관다발계의 안쪽에 제한되었으며 관다발계의 외부에 위치한 피층과 표피에서는 강하게 발현된 식물에서도 GUS 활성이 나타나지 않았다. GUS 활성은 어린 조직에서 특이적으로 발현되었으며 상처에 의해서 신속하게 증가하였다. RCaM-2 promoter는 세포분열이 왕성한 어린조직이나 생장점에서 강하게 발현되며 mechanical 신호에 의해서 현저히 유도되었다. 이러한 결과는 RCaM-2 유전자의 5'-flanking 영역이 상처에 의해서 유도되는 발현을 조절하는 것으로 추정된다.

• Plant Breeding and Biotechnology
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• 제2권3호
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• pp.289-300
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• 2014

A tissue-specific and developmentally expressed gene was isolated from Chinese cabbage (Brassica rapa L. ssp. pekinensis), designated BrREF (B. rapa Rubber elongation factor). BrREF transcripts were expressed at high levels in seedlings and at low levels in flower buds and roots. To study the activity of this promoter, the 2.2 kb upstream sequence of BrREF gene was fused to a β-glucuronidase (GUS) reporter gene and was introduced into Arabidopsis thaliana and B. napus by Agrobacterium-mediated transformation. Strong expression of GUS driven by the BrREF promoter was detected in the cotyledons and hypocotyls of transgenic plant seedlings, but GUS expression was weak in roots, excluding the root tips. GUS expression in the cotyledons and hypocotyls decreased dramatically as the seedlings matured and was not detected in the tissues of mature plants. During floral development, GUS expression was observed in immature anthers. These findings suggest that the BrREF promoter can modulate the tissue-specific and developmental expression of gene at the early stages of growth and development.