Promoters for milk proteins have been used far producing transgenic animals due to their temporal and spatial expression patterns. -casein, a calcium-sensitive casein, is a major milk protein that corresponds ca. 30 per cent of total milk protein. Expression of -casein is controlled by lactogenic hormones such as prolactin (PRL), composite response elements (CoREs) and transcription factors. CoREs are clusters of transcription factor binding sites containing both positive and negative regulatory elements. -casein gene promoter contains various regions (CoREs) for gene transcription. We analyzed the promoter region by mutagenesis using exonuclease III and linker-scanning. Transcription control elements usually are positioned in 5'-flanking region of the gene. However, in some cases, these elements are located in other regions such as intron 1. The nucleotide sequences of -casein promote. region has been reported (E12614). However, the properties of the promoter is not yet clear. In this study, we plan to investigate the properties of cis-regulating elements of porcine -casein by mutation analysis and expression analysis using dual-luciferase repoter assay system.

Anthocyanins are the major pigments contributing to flower coloration. A 1584 bp 5' upstream sequence of ALCHS2 gene was isolated from Acapulco lily (Lilium Oriental hybrid cv. Acapulco). Computer-based analyses (GeneScan, AtPAN) predicted a CAATBOX1 and putative transcription factor-binding sites, including tissue-specific elements. When gALCHS7 promoter–gus fusion was introduced to petunia ('Dream Red'), all ten putative transgenic plants showed localized GUS activity in the anther, but five of them also showed weak GUS activity in the ovule. No distinctive signal in the leaf and petal was detected in the same stage. To clearly determine the operation of the promotor region, anther and ovule tissues of transgenic line 6 were fixed in paraffin for dark-field analysis. At 1 cm length of floral bud, a GUS signal was not observed in the anther, but weak expression was observed in the ovule. Before anthesis, GUS protein was highly expressed in the pollen, endothecium, and epidermis. Fluorometric GUS assays of individual organs taken from four transgenic plants demonstrated that all lines showed high GUS activity in the anther compared to 35S CaMV promoter (pBI1 121), except line 34. Using the truncated promoters by cis-acting elements, we found that minimal region (gALCHS7-7, 270 bp) displayed GUS expression only in the anther, though at weaker activity than in the original promoter.

Zoysiagrass are damaged by fungi diseases such as large patch, dollar spot, pythium blight and brown patch. Large patch is one of the major diseases caused by Rhizoctonia solani AG2-2 on zoysiagrass fields e.g. golf courses. Plant chitinases have been known PR (Pathogen related)-protein. In this study, we isolated two chitinase genes (Zjchi1 and Zjchi2) from zoysiagrass. Antifungal activity analysis revealed that Zjchi2 protein inhibited mycelium extension of fungi. A further study, we cloned 5` upstream region from two chitinase genes for investigating transcription regulatory mechanism that inducing of two chitinase genes dependent R. solani. -818 bp and -799 of upstream region from Zjchi1 and Zjchi2 successfully isolated using in vitro LA (Long and Accurate) PCR system. And then, we generated promoter-GUS reporter constructs with deletion construct based on W-boxes. Constructs were introduced into Arabidopsis thaliana by Agrobacterium-mediated transformation for stable expression of GUS reporter gene.

클레마티스의 번식은 종자 및 삽목번식으로 하는데, 삽목 성공 률이 낮아 번식에 어려움이 있다. 이에 효과적인 번식법으로 삽목 번식 방법을 구명하고자 클레마티스 삽수를 채취하여 유리온실에 서 본 실험을 수행하였다. 삽수길이는 1마디와 2마디 길이로 하였 으며, 배지는 단용배지로 피트모스, 펄라이트, 버미큘라이트, 발근 제는 IBA 4000mg·L-1 3초 침지, IBA 500mg·L-1 30분 침지와 루 톤을 처리하였다. 클레마티스의 발근기간은 약 10주가 소요되었 다. 클레마티스의 삽수 마디길이를 2마디보다 1마디로 하였을 경 우 발근율이 약 43% 증가하였다. 1마디 삽수의 경우 발근제 루톤 분의 처리 시 발근율은 대조구보다 약 7% 더 향상되었다. 따라서, 삽수 양이나 삽목 후 관리를 고려할 때, 1마디 삽수를 이용하는 것 이 효과적인 것으로 보인다. 배지와 발근제에 따른 발근율은 버미 큘라이트와 IBA 4,000mg·L-1 처리에서 96%로 가장 높았으며, 뿌 리수와 뿌리 길이도 IBA 4,000mg·L-1 및 루톤 처리에서 우수하였 다. 즉, 1마디 삽수를 버미큘라이트 배지에 삽목한 경우 발근율은 평균 약 80-83%로 피트모스와 펄라이트보다 약 25-29%, 13-39% 더 높았으며, 버미큘라이트 배지에 IBA 4,000mg·L-1 침지처리 시 발근율이 89-96%로 9-13%가 더 증가되어 가장 효과적이었다

Carotenoid isomerase (CRTISO) catalyzes the isomerization of prolycopene to all-trans-lycopene in the carotenoid biosynthetic pathway. We isolated a full-length promoter region of CuCRTISO from Citrus unshiu. We determined if the promoter encoded organ-specific or developmental-specific expression, and identified possible cis-acting promoter elements. The full-length promoter and two truncated versions were fused to the β-glucuronidase (GUS) gene and transformed into Arabidopsis thaliana. Transgenic lines expressing the full-length promoter (pCiso-Prom1) and truncated promoters (pCiso-Prom2 and pCiso-Prom3) showed the same developmental and organ-specific activity. GUS expression was detected in the cotyledon and root at 5 and 10 days after germination, mature leaf, and anther. The CuCRTISO promoter contained several cis-acting elements involved in hormonal and environmental stress. Drought stress or abscisic acid treatment did not induce GUS expression in any transgenic lines. Heat stress induced GUS expression in the pCiso-Prom1 line; this promoter construct contains the heat-stress responsive element (HSE). Ethylene and cold-stress treatments induced GUS expression only in the pCiso-Prom3 line, although all transgenic lines contained the same cis-acting ethylene and low-temperature response elements. which could indicate the existence of unknown repressor element(s) in the CuCRTISO promoter. These studies indicate that CuCRTISO promoter activity is regulated in a developmental and organ-specific manner that responds to heat, cold, and ethylene. These results provide new insights into the role of cis-acting element(s) in CuCRTISO promoter activity. (This research was supported by the Basic Science Research Program of the National Research Foundation of Korea (NRF) funded by the Ministry of Education (NRF-2010-0007627 and 2009-0094059), and by Golden Seed Project, Ministry of Agriculture, Food and Rural Affairs (MAFRA), Ministry of Oceans and Fisheries (MOF), Rural Development Administration (RDA) and Korea Forest Service (KFS), Republic of Korea)

In plants, the Dof (DNA binding with One Finger) proteins are plant-specific transcription factors with a particular class of zinc-finger DNA-binding domain. The Dof genes have been predicted 30 different Dof genes in the rice Oryza sativa genome by phylogenetic analysis. The mostly Dof proteins contain a conserved region of 50 amino acids with a C2-C2 zinc finger motifs that binds a cis-regulatory element sequence 5’-T/AAAAG-3’. We found that a member of the DOF transcription factor family, Dof1 gene of rice, was expressed to wound from Ds insertion mutant population. Sequencing of the flanking regions of the transposon insertion site indicated that the gene-trap had been inserted near the front of the second exon of OsDof1 gene in chromosome 7. Genomic southern analysis revealed that mutant line contained a single copy of Ds gene trap. The Ds tagged rice mutant line, OsDof1::Ds, wound-inducible GUS expression was identified. To analyze the cis-acting elements, we constructed fusion genes with the OsDof1 promoter fused to the β-glucuronidase (GUS) reporter gene and transformed Arabidopsis and rice plants with these constructs. Wound-induced GUS expression was observed in the leaves of transgenic OsDof1::GUS rice and Arabidospsis plants. These results showed that, OsDof1 protein might be involved in stress responses and growth regulation in plant, might plays a role as a transcription regulator in stress response signal transduction pathways of plant.

나비류 추출물은 전통적으로 다양한 활성을 보유하여 의약용으로 사용되어졌다. 5종의 나비(산제비나비[Papilio maackii], 호랑나비[Papilio xuthus], 배추흰나비[Pieris rapae], 남방호랑나비[Eurema hecabe], 왕오색나비[Sasakia charonda])를 이용하여 물, dimethly sulfoxide (DMSO), ehtanol 및 methanol로 추출한 추출물로 항산화 활성 및 Cox-2 promote

We developed a novel dicistronic system for the expression of target cDNA sequences in the milk of transgenic animals using goat beta-casein/hGH fusion construct, pGbc5.5hGH (Lee, 2006) and internal ribosome entry site (IRES) sequences of encephalomyocarditis virus (EMCV). Granulocyte colony-stimulating factor (hG-CSF) cDNA was linked to 3' untranslated region of hGH gene in the pGbc5.5hGH via EMCV IRES sequences. Transgenic mice were generated by microinjection and transgene expression was examined in the milk and mammary gland of transgenic mice at 10 days of lactation. Northern blot analysis showed that hGH gene and hG-CSF cDNA were transcribed as a single dicistronic mRNA. The hG-CSF and hGH proteins were independently translated from the dicistronic mRNA and secreted into the milk of transgenic mice. The highest concentration of hG-CSF and hGH in the milk of transgenic mice were and , respectively. In contrast, another hG-CSF expression cassette, in which hG-CSF genomic sequences were inserted into a commercial milk-specific expression vector (pBC1), generated a lower level () of hG-CSF expression in the milk of transgenic mice. These results demonstrated that the novel pGbc5.5hGH-based dicistronic construct could be useful for an efficient cDNA expression in the milk of transgenic animals.