• Journal of Microbiology and Biotechnology
• /
• 제13권5호
• /
• pp.819-822
• /
• 2003

Polyketides are an extensive class of secondary metabolites with diverse molecular structures and biological activities. A plasmid-based minimal polyketide synthase (PKS) expression cassette was constructed using a subset of actinorhodin (act) biosynthetic genes (actI-orfl, actI-orf2, actI-orf3, actIII, actⅦ, and actIV) from Streptomyces coelicolor, which specify the construction of an orange-fluorescent anthraquinone product aloesaponarin II, a type II polyketide compound derived from one acetyl coenzyme A and 7 malonyl coenzyme A extender units. This system was designed as an indicator pathway in S. parvulus to generate a hybrid type II polyketide compound via gene-specific replacement. The act ${\beta}-ketoacyl$ synthase unit (actI-orfl and actI-orf2) in the expression cassette was specifically replaced with oxytetracycline ${\beta}-ketoacyl$ synthase otcY-orfl and otcY-orf2). This plasmid-based hybrid PKS cassette generated a novel orange-fluorescent compound structurally different from aloesaponarin II in both S. lividans and S. parvulus. In addition, several additional distinctive blue-fluorescent compounds were detected, when this hybrid PKS cassette was expressed in S. coelicolor B78 (actI-orf2 mutant), implying that the expression of plasmid-based hybrid PKS cassette in Streptomyces species should be an efficient way of generating hybrid type II polyketide compounds.

• 비교문화연구
• /
• 제47권
• /
• pp.217-240
• /
• 2017

본고는 피동 표현에서의 한국어 행위주 표지 '-에게(한테)', '-에, -로'와 중국어 피동 표지 '피(被)[$b{\grave{e}}i$]/양(?)[$r{\grave{a}}ng$]/규(叫)[$ji{\grave{a}}o$]/급(?)[$g{\check{e}}i$]'를 연구 대상으로 삼아 그 분포양상 및 특징을 살피고 이들 형태를 비교 대조를 통하여 그 대응 관계를 면밀히 고찰해 보고자 하였다. 대조 분석 시 두 언어의 유형적 특징, 피동 표현에서의 행위주, 피동주에 대한 선택 제약, 그리고 '받다'류 피동 표현에서 행위주(피동) 표지의 사용 제약과 같은 3가지 측면에 중점을 두었다. 본 대조 분석을 통해 확인한바, 한 중 피동 표현에서 한국어 행위주 표지 '-에게(한테)', '-에, -로'와 중국어 피동 표지 '피(被)/양(?)/규(叫)/급(?)'는 각각 행위주와 결합하여 부사어 역할을 하고 있는 공통점에도 불구하고 용법에 있어 차이점을 보였다. 먼저 두 언어 유형적 특징에 따라 피동 표현에서 행위주와의 결합 방식이 각각 달리 나타남을 확인하였다. 그리고 한국어 행위주 표지는 오로지 조사 역할을 하여 '행위주 유정성 유무'에 대한 제약만 받는 반면, 중국어 피동 표지 '피(被)/양(?)/규(叫)/급(?)'는 각각 문법화 정도에 따라 행위주 유무뿐만 아니라 피동주 유정성 유무 그리고 문장에 나타나는 어휘의 의미에 따른 제약 등을 보인다. 특히 한국어 '받다'류 피동 표현에서 한국어의 행위주 표지 '에게(한테), -에, -로'는 그대로 사용되지만 대응하는 중국어에서는 피동 표지 사용에 여러 제약이 있음을 확인하였다.

• Journal of Plant Biotechnology
• /
• 제40권1호
• /
• pp.27-36
• /
• 2013

We isolated and functionally characterized a Dendrobium Actin1 (DmACT1) promoter that drives strong gene expression in the orchid genus Dendrobium. A genomic fragment containing the region 3227 bp upstream of the coding region of DmACT1 was obtained by inverse PCR. Detailed comparison of the full-length cDNA and genomic sequences revealed that DmACT1 has a 1374 bp first intron in the 5' UTR. However, the 5' flanking sequences upstream of the coding region showed no obvious sequence similarities compared to those of known promoters, including plant actin promoters. Serial deletion constructs of the 5' flanking region from the translation initiation codon were fused to the coding sequence of a GUS/luciferase fusion reporter to identify the regulatory elements necessary for promoter activity. Transient assays in the flowers of Dendrobium revealed that the 5' UTR-intron greatly enhanced promoter activity. Moreover, the DmACT1 promoter with its 5' UTR-intron yielded approximately 10-fold higher reporter activity than the rice Act1 promoter-intron. Our data suggest that the DmACT1 promoter with its 5' UTR-intron is a useful tool for strong expression of transgenes in Dendrobium orchids.

• Journal of Plant Biology
• /
• 제38권4호
• /
• pp.365-371
• /
• 1995

Binary vectors, pBI-ActR1, pBI-ActF1 and pBSH-ActR1, were constructed using pGA642, the replication origin of pTi12 and the rice actin promoter. The sizes of pBI-ActR1, pBI-ActF1 and pBSH-ActR1 were 12.9 kb, 13.2 kb and 11.95 kb, respectively. These vectors containing a rice actin promoter followed by a GUS structural gene could induce stronly the expression of GUS gene in transformed rice cells. Rice explants from 3-4 day old seedlings after germinatin were cocultured with A. tumefaceins harboring pBI-ActR1, pBI-ActF1 or pBSH-ActR1, and then GUS expression in the explants was assayed. Transformation of rice explants by these binary vectors was tissue-specific, such that the meristematic regions of shoot apex, root and hypocotyl were transformed by these binary vectors.

• Journal of Plant Biology
• /
• 제37권3호
• /
• pp.371-380
• /
• 1994

To determine the patterns and the levels of expression of the cauliflower mosaic virus (CaMV 35S) promoter and the rice actin 1 (Act1) promoter in rice, transgenic rice plants containing CaMV 35S-$\beta$-glucuronidase (GUS) and Act1-GUS constructs were generated and examined by fluorometric and histochemical analyses. The fluorometric analysis of stably transformed calluses showed that the activity of the rice Act1 promoter was stronger than that of the CaMV 35S promoter in rice cells. In a histochemcial study of the transgenic rices, it was shown that the GUS activity directed by the CaMV 35S promoter was localized mainly in parenchymal cells of vascular tissues of leaves and roots and mesophyll cells of leaves. These results are similar to those of potato, a dicot plant. In contrast, rice plant transformed with Act1-GUS fusion construct revealed strong GUS activity in parenchymal cells of vascular tissue, mesophyll cells, epidermal cells, bulliform cells, guard subsidiary cells of leaves and most cells of the root, suggesting that the rice Act1 promoter is more constitutive than the CaMV 35S promoter. It was also confirmed that in both types of transgenic rice little or no staining was localized in metaxylen tracheary elements of vascular tissue from leaves or roots. These results indicate that the rice Act1 promoter can be utilized more successfully for expression of a variety of foreign gene in rice than the CaMV 35S promoter.

• 한국발생생물학회지:발생과생식
• /
• 제26권4호
• /
• pp.165-174
• /
• 2022

Three proteins [myosin heavy chain (MHC), filamin-C fragment (FIL-C), and actin 2 (ACT2)] were identified in adductor muscle from diploid and triploid Pacific oysters (Crassostrea gigas) and the relationship between the condition index (CI) and mRNA expression of these genes was investigated, together with the mRNA expression of molluscan insulin-related peptide (MIP), C. gigas insulin receptor-related receptor (CIR), and insulin-like growth factor binding protein complex acid labile subunit (IGFBP-ALS). Monthly changes in the CI were similar to the changes in the tissue weight rate in both groups. ACT2 and MHC mRNA expression was statistically higher in the triploid than the diploid, while FIL-C mRNA expression was significantly higher in the diploid (p<0.05). The MIP, CIR, and IGFBP-ALS mRNA expression of the diploid oysters were all significantly higher in July than in other months (p<0.05). The MIP, CIR, and IGFBP-ALS mRNA expression in the triploid oysters was high in July, but there were no significant differences (p>0.05). Changes in the expression levels of the genes investigated in this study could be used as intrinsic indicators of the annual growth, maturity, and spawning period of cultured diploid and triploid C. gigas in Tongyeong, Korea.

• 식물병연구
• /
• 제26권3호
• /
• pp.144-158
• /
• 2020

정량적역전사중합효소연쇄반응(quantitative reverse transcription PCR)은 정확하고 민감한 방법으로 유전자 발현분석에 사용된다. 사과 식물에서 유전자 발현 변화의 정량적 변화를 분석하기 위해, 사과 잎검은점 바이러스(Apple stem grooving virus, ASGV)에 의한 감염 동안 발현의 안정성에 대해 10개 참조유전자들(ACT, CKL, EF-1α, GAPDH, MDH, PDI, THF, UBC, UBC10 및 WD40)을 평가하였다. AGSV 감염 또는 저온 처리된 사과 식물에서의 10개 참조유전자 발현의 안정은 5가지 프로그램을 사용하여 분석하였다. ASGV 감염 사과식물의 잎 조직에서는 CKL>THFs>GAPDH>ACT 순서로 가장 안정한 유전자로 분석되었으며 WD40CKL>UBC10이고 가장 안정하지 않은 유전자는 ACT

• Archives of Pharmacal Research
• /
• 제26권1호
• /
• pp.53-57
• /
• 2003

We have found that ginsenoside Rc and Re induce c-fos in MCF-7 human breast carcinoma cells at both the mRNA and protein levels. However, neither ginsenoside activated the expression of reporter gene under the control of AP-1/TPA response elements. We have also examined the possibility that ginsenoside Rc and Re act by binding to intracellular steroid hormone receptors that act as transcriptional factors in the nucleus in inducing c-fos mRNA in MCF7 human breast carcinoma cells. However, ginsenoside Rc and Re did not bind to glucocorticoid, androgen, estrogen, or retinoic acid receptors as examined by the transcription activation of the luciferase reporter genes in CV-1 cells that were transiently transfected with the corresponding steroid hormone receptors and hormone responsive luciferase reporter plasmids. These data demonstrate that ginsenoside Rc and Re act via other transcription factors and not via estrogen receptor in c-Fos expression.

• 한국식품저장유통학회지
• /
• 제15권1호
• /
• pp.84-87
• /
• 2008

유전자 변형 작물은 생산성 측면에서 많은 장점이 있지만 이를 섭취할 경우 잠재적인 위험 요소들에 의해 많은 문제가 대두대고 있다. 본 연구는 저항성유전자를 이입한 배추에서 Profillin, Tubulin-${\alpha}$ (Tub-${\alpha}1$), Heat-shock protein (Bchsp 17.6) and Ubiquitin conjugating enzyme (UBE)의 발현과 이를 30일간 섭취한 마우스에서 ${\beta}$-actin(${\beta}$-act), ${\beta}$-2-microglobulin (B2m), Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and ${\beta}$-glucuronidase (Gus)의 발현 정도를 RT-PCR을 통해 알아보았다. 실험 결과 저항성유전자를 이입한 배추와 그렇지 않은 배추의 유전자 발현 패턴은 큰 차이를 보이지 앓았으며, 이를 섭취한 마우스 장기에서도 발현에 따른 큰 차이는 나타나지 않았다.

• Clinical and Experimental Reproductive Medicine
• /
• 제26권3호
• /
• pp.383-387
• /
• 1999

Objective: There is increasing evidence for the expression of rat in gene in several extrapituitary sites including testis and ovary. We also have demonstrated that the local LH expression in the rat epididymis and uterus, the major accessory sex organs in male and female reproductive system, respectively. Design: The present study was undertaken to elucidate whether the gene for LH receptor is expressed in rat uterus and whether the expressions of uterine LH and its receptor are differentially regulated during estrous cycle. Presence of the transcripts for rat LH receptor in the rat uterine tissue were confirmed by touchdown reverse transcription-polymerase chain reaction (RT-PCR). Results: In $LH{\beta}$ semi-quantitative RT-PCR, the highest expression level was shown in estrus stage. The level of ill receptor transcripts was also fluctuated during estrous cycle. In ovariectomized rats (OVX + Oil), the expressions of both uterine LH and LH-R were markedly reduced when compared to those from normal rats. Supplement with estradiol $17{\beta}$ to the ovariectomized rats (OVX + $E_2$) restored the expression levels of LH and its receptor to the levels in uteri from normal rats. Conclusion: Our findings indicated that 1) LH and its receptor gene are expressed in the rat uterus from cycling rats, 2) the expression of uterine LH and its receptor is mainly, if not all, under the control of ovarian sex steroid(s). These results suggested that the uterine LH may act as a local regulator with auto and/or paracrine manner, though the posibility that the pituitary LH may act directly on the regulation of uterine functions could not be discarded.