Bacteriophage T7 gene 2.5 protein, a single-stranded DNA binding protein, has been implicated in T7 DNA replication, recombination, and repair. Purified gene 2.5 protein has been shown to interact with the phage encoded gene 5 protein (DNA polymerase) and gene 4 proteins (helicase and primase) and stimulates their activities. Genetic analysis of T7 phage defective in gene 2.5 shows that the gene 2.5 protein is essential for T7 DNA replication and growth. T7 phage that contain null mutants of gene 2.5 were constructed by homologous recombination. These mutant phage $(T7\Delta2.5)$ cannot grow in Escherichia coli. After infection of E. coli with $T7\Delta2.5$, host DNA synthesis is shut off, and $T7\Delta2.5$ DNA synthesis is reduced to less than $1\%$ of wild-type phage DNA synthesis (Kim and Richardson, 1993, Proc. Natl. Aca. Sci. USA, 90, 10173-10177). A truncated gene 2.5 protein $(GP2.5-\Delta21C)$ deleted the 21 carboxyl terminal amino acids was constructed by in vitro mutagenesis. $GP2.5-\Delta21C$ cannot substitute for wild-type gene 2.5 protein in vivo; the phage are not viable and exhibit less than $1\%$ of the DNA synthesis observed in wild-type phage-infected cells. $GP2.5-\Delta21C$ has been purified to apparent homogeneity from cells overexpressing its cloned gene. Purified $GP2.5-\Delta21C$ does not physically into「act with T1 gene 4 protein as measured by affinity chromatography and immunoblot analysis. The mutant protein cannot stimulate T7 gene 4 protein activity on RNA-primed DNA synthesis and primer synthesis. These results suggest that C-terminal domain of gene 2.5 protein is essential for protein-protein interactions.
Choi, Eun Hwa;Kim, Hee Sup;Chanock, Stephen J.;Lee, Hoan Jong
Clinical and Experimental Pediatrics
/
v.48
no.5
/
pp.495-499
/
2005
Purpose : Interleukin-4(IL-4) is a critical component of the Th2 cytokine pathway and contributes to severity of respiratory syncytial virus(RSV) bronchiolitis. Previous studies observed an association between severe RSV bronchiolitis in Korean children with a common haplotype of the IL4 promoter. This study was performed to investigate functional differences of the variant IL4 promoter haplotypes. Methods : Genomic DNA was obtained from 20 children from 6 to 48 months of age in the Department of Pediatrics, Seoul National University Bundang Hospital. The IL4 promoter spanning an 1.2 kb region was amplified and haplotype was determined by cloning and the PHASE reconstruction. Transcriptional activity of Jurkat T cells which were transfected with each IL4 haplotype were analyzed by use of luciferase assay. Results : Three haplotypes of the IL4 promoter have been identified with the frequency of GCC(7 percent), TCC(17 percent), and TTT(76 percent). The TTT haplotype demonstrated the highest luciferase values in both unstimulated and PMA-stimulated Jurkat T cells. Increases in transcriptional activity compared to GCC have been shown in TTT(5.3 fold higher) followed by TCC(4.2 fold higher) in unstimulated Jurkat T cells. Conclusion : We provided evidence that increased transcriptional activity of the TTT haplotype of the IL4 promoter, which has previously been over-represented in Korean children with severe RSV bronchiolitis. Therefore, IL-4 could play a potential role in the pathogenesis of RSV infection, possibly via an altered transcriptional activity of the different IL4 haplotypes.
Disruptive expression patterns of the circadian clock genes are highly associated with many human diseases, including cancer. Cell cycle and proliferation is linked to a circadian rhythm; therefore, abnormal clock gene expression could result in tumorigenesis and malignant development. The molecular network of the circadian clock is based on transcriptional and translational feedback loops orchestrated by a variety of clock activators and clock repressors. The expression of 10~15% of the genome is controlled by the overall balance of circadian oscillation. Among the many clock genes, Period 1 (Per1) and Period 2 (Per2) are clock repressor genes that play an important role in the regulation of normal physiological rhythms. It has been reported that PER1 and PER2 are involved in the expression of cell cycle regulators including cyclins, cyclin-dependent kinases (CDKs), and CDK inhibitors. In addition, correlation of the down-regulation of PER1 and PER2 with development of many cancer types has been revealed. In this review, we focused on the molecular function of PER1 and PER2 in the circadian clock network and the transcriptional and translational targets of PER1 and PER2 involved in cell cycle and tumorigenesis. Moreover, we provide information suggesting that PER1 and PER2 could be promising therapeutic targets for cancer therapies and serve as potential prognostic markers for certain types of human cancers.
Hyo Seo Kang;Tae Hee Nam;Woo Ju Lee;Joon Sang Lee;Sangsu Shin
Korean Journal of Poultry Science
/
v.50
no.4
/
pp.261-266
/
2023
The skeletal muscles of livestock play a crucial role as protein sources for humans, and the consumption of poultry meat is steadily increasing worldwide. Numerous genes, including myogenic regulatory factors, are involved in myogenesis, and precise regulation of them is essential. In this study, genes specifically expressed in muscles were selected, and their promoters were cloned and analyzed. The analysis of gene expression in various tissues of animals revealed that many genes exhibited specific expression patterns in skeletal muscles, with TNNT3, TNNC2, and MYF6 genes showing similar patterns in poultry. The promoter regions of three genes were amplified by polymerase chain reaction to sizes of 1.2 kb, 1.03 kb, and 1.43 kb, respectively. These fragments were then inserted at the front of the enhanced green fluorescent protein gene in vectors. It was confirmed that the sequences of three promoters closely matched the chicken genome sequences. Upon introducing vectors with each promoter into QM7 quail muscle cells, all three promoters successfully induced the expression of the green fluorescent protein. The brightness of the green fluorescence in each promoter was approximately seven times dimmer compared to the control, CMV-IE promoter. It is predicted that more than 230 transcription factors can bind to each promoter, especially various transcription factors expressed in muscles, including myogenic regulatory factors such as MYF5, MYOD, and MYOG. These promoters can be valuable for studying gene expression in poultry muscle cells, and further research is needed to precisely investigate the regulatory region of gene expression in promoters.
A new rice variety 'Jinbo' is a japonica rice (Oryza sativa L.) with good eating quality, lodging tolerance, and resistance to rice stripe virus (RSV) and bacterial blight disease (BB). It was developed by the rice breeding team of Yeongdeog Substation, National Institute of Crop Science (NICS), RDA in 2009. This variety was derived from a cross between 'Yeongdeog26' with good grain quality and wind tolerance and 'Koshihikari' with good eating quality in 1998 summer season. A promising line, YR21324-56-1-1, selected by pedigree breeding method, was designated as the name of 'Yeongdeog45' in 2005. After the local adaptability test was carried out at nine locations from 2006 to 2008, 'Yeongdeog45' was released as the name of 'Jinbo' in 2009. 'Jinbo' has short culm length as 74 cm and medium maturating growth duration. This variety is resistant to $K_1$, $K_2$, and $K_3$ races of bacterial blight and stripe virus and moderately resistant to leaf blast disease with durable resistance, and also it has tolerance to unfavorable environments such as cold and dried wind. 'Jinbo' has translucent and clear milled rice kernel without white core and white belly rice, and good eating quality as a result of panel test. The yield potential of 'Jinbo' in milled rice is about 5.65 MT/ha at ordinary fertilizer level in local adaptability test. This cultivar would be adaptable to middle plain, mid-west costal area, east-south coastal area, and south mid-mountainous area.
A new rice variety 'Haeoreumi' is a japonica rice (Oryza sativa L.) with lodging tolerance, resistance to rice stripe virus (RSV) and bacterial leaf blight (BLB), and high grain quality. It was developed by the rice breeding team of Yeongdeog Substation, National Institute of Crop Science (NICS), RDA in 2008. This variety was derived from a cross between 'Milyang165' with good grain quality and lodging resistance, and 'Haepyeongbyeo' with wind tolerance in winter season of 2000/2001. A promising line, YR22375-B-B-1, selected by pedigree breeding method, was designated as the name of 'Yeongdeog46' in 2005. 'Yeongdeog46' was released as the name of 'Haeoreumi' in 2008 after the local adaptability test that was carried out at nine locations from 2006 to 2008. 'Haeoreumi' has 74 cm short culm length as and medium maturating growth duration. This variety showed resistance to $K_1,\;K_2$, and $K_3$ races of bacterial blight, and stripe virus and moderate resistant to leaf blast disease with durable resistance, and also has tolerance to unfavorable environment such as cold, dry and cold salty wind. 'Haeoreumi' has translucent and clear milled rice kernel without white core and white belly rice, and good eating quality as a result of panel test. The yield potential of 'Haeoreumi' in milled rice is about 5.58MT/ha at ordinary fertilizer level of local adaptability test. This cultivar would be adaptable to Middle plain, mid-west costal area, and east-south coastal area.
Proceedings of the Plant Resources Society of Korea Conference
/
2018.10a
/
pp.26-27
/
2018
With entry into force of the Nagoya Protocol to promote the fair sharing of the benefits of accessing and utilizing genetic resources, much support has been given to research on the development of biomaterials and products using domestic natural resources. Conservation and resource-saving of native species became very important through Nagoya Protocol enactment. The trend of cosmetic industry has been shifing from use synthetic chemicals to natural biomaterials, due to the safety regulations on new materials, ban on animal experiments, and expansion of cosmeceuticals range. In addition, functional cosmetic range has been expanded from whitening, wrinkle improvement, and ultraviolet shielding, to hair loss, hair loss alleviation, acne relaxation, and moisturizing of atopic skin, thus causing the activation of research about field of efficacy evaluation on natural biomaterials and commercialization. Chungbuk province is fostering the bio industry as a key industry for regional economic growth. For this purpose, Osong Biotechnology Complex/Ochang Science Industrial Complex in middle area, Jecheon biovalley in northern region, and Chungju Enterprise city have been established, thus playing a pivotal role in Bio innovative cluster in Korea. In particular, it was established the osong cosmetics clinical research support center to develop the cosmetics industry in chungbuk, thereby supporting clinical trials, efficacy evaluations, overseas certification, and overseas market entry in order to advance into the global market. In addition, oriental plants such as astragalus propinquus, schisandra chinensis, eucommia, alpiniae oxyphyllae fructus and biancaea sappan are being actively studied as global cosmetic ingredients through the promotion of various national research and development projects using natural materials in chungbuk province. The chungbuk natural product industry is expected to grow further throughout cosmetics industry development in the future, as companies and research institutes are actively promoting the secure index of effective material in natural products and effective material commercialization.
Seo, Young-Jun;Cheong, Han Bin;An, Seok Min;Sin, Woo Cheol;Bae, Eun Joo;Yoon, Jong Hyung;Jeong, Hwal Rim;Lee, Hong Jin
Journal of The Korean Society of Inherited Metabolic disease
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v.18
no.3
/
pp.87-94
/
2018
We present the case of long-term observation of a patient with chronic kidney disease (CKD) caused by advanced focal segmental glomerulosclerosis (FSGS) resulting from underlying congenital chloride diarrhea (CLD). A 20-year-old woman was admitted for prolonged proteinuria despite conservative treatment for CLD. She was diagnosed with CLD and started taking KCl salt supplementation from the time of birth. Mild proteinuria was first found at 12 years of age, which progressed to moderate proteinuria at 16 years of age. At 16 years of age, CKD stage 2 with FSGS was diagnosed based on the initial assessment of the glomerular filtration rate (GFR) and kidney histology. On admission, we re-assessed her renal function, histology and genetic analysis. GFR had deteriorated to CKD stage 4 and renal histology revealed an advanced FSGS combined with tubulointerstitial fibrosis. A homozygous mutation in the SLC26A3 gene (c.2063-1G>T) was found by diagnostic exome sequencing and may have been inherited from both parents. CLD patients can be more vulnerable to renal injury, which may also cause progression of renal failure. Therefore, even if there is an early diagnosis and adequate salt supplementation, close monitoring of renal function and tailored treatment should be emphasized for renal protection and favorable CLD prognosis.
Salt stress is one of major environmental factors influencing plant growth and development. To identify salt tolerance determinants in higher plants, a large-scale screen was conducted with a bialaphos marker-based T-DNA insertional collection of Arabidopsis ecotype C24 mutants. One line for salt stress-sensitive mutant (referred to as ssm1) exhibited increased sensitivity to both ionic (NaCl) and nonionic (mannitol) osmotic stress in a root growth assay. This result suggests that ssm1 mutant is involved in ion homeostasis and osmotic compensation in plant. Molecular cloning of the genomic DNA flanking T-DNA insert of ssm1 mutant was achieved by mutant genomic DNA library screening. T-DNA insertion appeared in the first exon of an open reading frame on F3M18.7, which is the same as AtSYP61. SSM1 is SYP61/OSM1 that is a member of the SNARE superfamily of proteins required for vesicular/target membrane fusions and factor related to abiotic stress.
Kim, Ok-Soo;Kim, Yong-Man;Kim, Nam-Young;Lee, Eo-Jin;Jang, Min-Kyung;Lee, Dong-Geun;Lee, Sang-Hyeon
Journal of Life Science
/
v.17
no.2
s.82
/
pp.167-173
/
2007
To study the transcriptional mechanisms by which expression of the murine glial cell-derived neurotrophic factor inducible transcription factor (mGIF) gene is regulated, a murine genomic clone was iso-lated using a mGIF cDNA as probe. A 13-kb genomic fragment, which comprises 4-kb upstream of the transcription initiation site was sequenced. The promoter region lacks a TATA box and CAAT box, is rich in G+C content, and has multiple putative binding sites for the transcription factor Spl. The mGIF gene also has consensus sequences for AP2 binding sites. The transcriptional activity of five deletion mutants of a 2.1-kb fragment was analyzed by modulating transcription of the heterologous luciferase gene in the promoterless plasmid pGL2-Basic. All mutants showed significant transcriptional activity in the murine neuroblastoma cell line NB41A3. Transient expression assays suggested the presence of a positive regulator between -213 and -129 while a negative regulator was found in the region between -806 and -214. Relatively strong transcriptional activity was observed in neuronal NB41A3, glial C6 cells and hepatic HepG2, but very weak activity in skeletal muscle C2C12 cells. These findings confirm the tissue-specific activity of the mGIF promoter and suggest that this gene shares structural and functional similarities with the dopamine receptor genes that it regulates.
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