• 식물병연구
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• 제17권2호
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• pp.111-120
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• 2011

A bacterial artificial chromosome library of Pectobacterium carotovorum 35 was constructed to characterize the genome and to sequence its hrp region. The hrp cluster of P. carotovorum 35 consisted of 26 open reading frames in five operons. A promoter-based green fluorescent protein technology was used to identify the genes regulated by the alternative sigma factor, HrpL, in P. carotovorum 35. The majority of the selected clones contained the hrpJ operon promoter sequence, which harbors a hrp box, but no putative hrp boxes were detected within the promoter sequences of two other hrpL-regulated genes encoding for pectate lyase and large repetitive protein. Although the promoters of five other hrp operons also contained hrp boxes, their expression was not HrpL-dependent in the promoter-based selection in E. coli. However, transcriptional analysis showed that expression from all operons harboring hrp boxes, except for the hrpN operon, was reduced significantly in the hrpL mutant. The severity of soft-rot symptoms when the hrpL mutant was applied to the surface of tobacco leaves, mimicking natural infection, was greatly attenuated. These results indicate that the hrpL gene of P. carotovorum 35 may be involved in the development of soft-rot symptoms.

• BMB Reports
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• 제29권4호
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• pp.359-364
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• 1996

By both in vitro hydroxylamine mutagenesis of the wild type 3-phosphoglycerate kinase gene (PGK) promoter DNA and insertion of the leu2-d gene, we have created yeast expression vectors potentially useful for production of eukaryotic genes in yeast. The guanine (G) to adenine (A) change at the -3 position from the ATG start codon of the PGK promoter-based vector rendered a 6~7 times elevated expression of the adjacent eukaryotic gene, and insertion of the leu2-d gene in the vector containing the mutated PGK promoter further enhanced the expression of the gene. When expression of the AIDS virus HIV1-gagP17 gene in a lacZ fusion form was examined with this new vector, a 15 times higher level of expression than that from the original PGK promoter was observed. Northern and Southern analysis showed that this elevated expression is due to the production of a high copy number of mRNA by leu2-d gene functioning and by efficient translation of the produced mRNA. Thus, the vector that contained the A at the -3 position from the ATG start codon in the promoter region and the leu2-d gene shows increased expression capability and will be potentially useful for production of eukaryotic genes in yeast.

• 한국발생생물학회지:발생과생식
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• 제15권1호
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• pp.25-30
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• 2011

Embryonic stem (ES) cells can self-renew maintaining the undifferentiated state. Self renewal requires many factors such as Oct4, Sox2, FoxD3, and Nanog. NF-${\kappa}B$ is a transcription factor involved in many biological activities. Expression and activity of NF-${\kappa}B$ increase upon differentiation of ES cells. Reportedly, Nanog protein directly binds to NF-${\kappa}B$ protein and inhibits its activity in ES cells. Here, we found a potential binding site of NF-${\kappa}B$ in the human Nanog promoter and postulated that NF-${\kappa}B$ protein may regulate expression of the Nanog gene. We used human embryonic carcinoma (EC) cells as a model system of ES cells and confirmed decrease of Nanog and increase of NF-${\kappa}B$ upon differentiation induced by retinoic acid. Although deletion analysis on the DNA fragment including NF-${\kappa}B$ binding site suggested involvement of NF-${\kappa}B$ in the negative regulation of the promoter, site-directed mutation of NF-${\kappa}B$ binding site had no effect on the Nanog promoter activity. Furthermore, no direct association of NF-${\kappa}B$ with the Nanog promoter was detected during differentiation. Therefore, we conclude that NF-${\kappa}B$ protein may not be involved in transcriptional regulation of Nanog gene expression in EC cells and possibly in ES cells.

• 생명과학회지
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• 제9권1호
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• pp.50-57
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• 1999

STAT들은 다양한 cytokine이나 성장호르몬 등에 의해 활성화되어 핵으로 빠르게 이동되어 유전자 발현을 조절한다. 우리는 D-raf 유전자의 5' 발현조절영역에서 STAT결합부위 염기서열을 발견하였다. 본 연구는 D-raf 유전자발현조절에 있어서 STAT결합부위의 역할을 형질전환 초파리를 사용하여 조사하였다. STAT결합부위에 염기치환돌연변이를 도입시킨 D-raf promoter부분을 P인자 벡터의 IacZ유전자에 융합시킴으로써 리포터 플라스미드 pDraf-STATmut-lacZ를 제작하였다. 이 리포터 플라스미드를 이용하여 얻은 Draf-STATmut-lacZ형질전환 초파리의 lacZ발현을 발생단계별, 조직별로 조사한 결과, 거의 모든 발생단계에서 정상형 Draf-lacZ 형질전환 초파리의 lacZ발현에 비해 크게 감소하였다. 본 연구결과들은 STAT결합배열 부위가 D-raf유전자발현조절에 있어서 중요한 역할을 함을 개체수준에서 증명하고 있다.

• 농업과학연구
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• 제38권4호
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• pp.673-680
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• 2011

The aim of this study was to identify the polymorphism on fatty acid binding protein (FABP4) gene promoter region and its association with carcass traits in Hanwoo. We performed PCR-direct sequencing of FABP4 promoter region to identify single nucleotide polymorphism (SNPs) using unrelated 24 Hanwoo bulls. Four SNPs (-298A>G, -472A>G, -887A>G, -862A>G) were detected in the promoter region and genotyped on 583 Hanwoo steers. A linear mixed model revealed an association of three SNPs (-298A>G, -472A>G and -862A>G) with carcass weight and marbling score in dominance model (P<0.05). The animals with AA genotypes for the three SNPs were heavier carcass weight (5 kg) than animals with GG genotypes in the statistical analysis. For the marbling score, the AA genotype was lower effect of marbling score (0.21) than GG genotypes. In conclusion, this study indicates an important role for three SNPs detected in promoter region of FABP4 in determining carcass weight and marbling score in Hanwoo.

• Animal cells and systems
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• 제2권2호
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• pp.287-295
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• 1998

The Saccharomyces cerevisiae MGMT gene encodes a O6-methylguanine DNA methyltransferase that protects cells from mutation or death by DNA alkylating agents. Using an in vitro transcription system, we analyzed its promoter region to find regulatory elements for transcription initiation. DNase I footprinting and a transcription assay showed that a functional TATA box, 5'-TGATATAGCA-3', is located in the region spanning from -25 to -34. We also found one upstream repressing sequence (URS), -333 to -213, by promoter deletion and competition analysis. Gel mobility shift assays and Southwestern blot analysis using URS region indicate specific complex formations. These results indicate that several cis-acting and trans-acting elements might be involved in the transcriptional regulation of the S. cerevisiae MGMT gene.

• Asian Pacific Journal of Cancer Prevention
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• 제15권19호
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• pp.8451-8454
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• 2014

Objectives: Epidemiological studies have shown that molecular mechanisms underlying the development of lung cancers differ between smokers and unsmokers. Aberrant promoter methylation in some tumor suppressor genes is frequent in lung tumors from smokers but rare in those from non-smokers. Recently, many studies have investigated the association between cigarette smoking and RASSF1A gene promoter hypermethylation in lung cancer patients, but a unanimous conclusion could not be reached. We therefore performed this meta-analysis to derive a more precise estimation of any association. Study Design: An electronic search of PubMed and Chinese Biomedicine databases was conducted to select studies. A total of 19 case-control studies were chosen, and odds ratios (ORs) with confidence intervals (CIs) were used to assess the strength of associations. Results: The case-control studies covered 2, 287 lung cancer patients: 63.4%(1449) of the patients were smokers, 36.6% (838) were unsmokers. The overall results suggested that smokers with lung cancer had a 1.297-fold (95% CI: 1.066~1.580, p=0.010, p=0.087) higher risk for RASSF1A gene hypermethylation than the non-smokers. In the stratified analysis, an increased risk of RASSF1A gene hypermethylation in smokers than in non-smokers was found in Asian (OR=1.481, 95%CI: 1.179~1.861, p=0.001, p=0.186). Conclusions: This meta-analysis supports the idea that RASSF1A gene hypermethylation is associated with cigarette smoking-induced lung cancer.

• 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.

• 한국잠사곤충학회지
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• 제52권1호
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• pp.39-44
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• 2014

효율적인 형질전환 누에 시스템 구축을 위해서는 새로운 전이인자의 개발과 함께 선발을 위한 마커 유전자 및 transposase 발현을 효과적으로 조절할 수 있는 다양한 유전자 프로모터 개발이 필수적이다. 이와 관련하여 선행연구를 통해 누에 후부실샘으로부터 고발현하는 RNA binding protein-1 homologue(RBP-1) 유전자를 선발한 바 있다. 본 연구에서는 RBP-1유전자의 누에 발육시기별 및 유충 조직별 발현양상을 Northen blot hybridization 방법으로 분석한 결과, RBP-1 유전자는 유충기로부터 번데기 후기까지의 전기간에 걸쳐 발현하였으며, 두부, 표피, 중장, 지방체 및 견사선 등 실험한 모든 유충 조직에서 고발현 하는 것으로 관찰되었다. 또한, 누에 게놈 유전자은행을 제작한 후 RBP-1 cDNA 유전자를 탐침으로 5'-UTR 영역을 클로닝하고 luciferase assay 방법으로 RBP-1 유전자 프로모터의 활성을 분석하였다. 실험 결과, RBP-1 cDNA를 탐침으로 RBP-1 유전자 ORF와 5'-UTR이 포함된 약 1,660 bp 영역의 게놈 유전자를 클로닝하였다. RBP-1 유전자 프로모터 활성검정을 위해 전사 개시점(+ 30)으로부터 상류의 -740 bp 영역을 PCR로 분리한 후 pGL3 basic vector에 도입하여 luciferase 활성 측정을 위한 전이벡터, pGL-RBP1를 제작하였다. 제작된 pGL-RBP1는 곤충 세포주(Sf9)에 transfection 한 후 luciferase 발현량을 측정한 결과, 기존의 BmA3 유전자 프로모터 대비 10% 가량 높은 발현 효율을 확인할 수 있었다.

• Asian-Australasian Journal of Animal Sciences
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• 제21권12호
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• pp.1689-1695
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• 2008

Leptin, the hormone product of the obese gene, is secreted predominately from white adipose tissue and regulates feed intake, energy metabolism and body composition. It has been considered a candidate gene for performance, carcass and meat quality traits in beef cattle. The objective of this study was to identify SNPs in the promoter region of the leptin gene and to evaluate the possible association of the SNP genotypes with carcass and meat quality traits in Korean cattle. We identified a total of 25 SNPs in the promoter region (1,208-3,049 bp upstream from the transcription start site) of the leptin gene, eleven (g.1508C>G, g.1540G>A, g.1545G>A, g.1551C>T, g.1746T>G, g.1798ins(G), g.1932del(T), g.1933del(T), g.1934del(T), g.1993C>T and g.2033C>T) of which have not been reported previously. Their sequences were deposited in GenBank database with accession number DQ202319. Genotyping of the SNPs located at positions g.2418C>G and g.2423G>A within the promoter region was performed by direct sequencing and PCR-SSCP method to investigate the effects of SNP genotypes on carcass and meat quality traits in Korean cattle. The SNP and SSCP genotypes from the two mutations of the leptin promoter were shown to be associated with the BF trait. The average BF value of animals with heterozygous SNP genotype was significantly greater than that of animals with the homozygous SNP genotypes for the g.2418C>G and g.2423G>A SNPs (p<0.05). Analysis of the combined genotype effect in both SNPs showed that animals with the AC SSCP genotype had higher BF value than animals with BB or AA SSCP genotypes (p<0.05). These results suggest that SNP of the leptin promoter region may be useful markers for selection of economic traits in Korean cattle.