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        Construction of fat1 Gene Expression Vector and Its Catalysis Efficiency in Bovine Fetal Fibroblast Cells

        Liu, Boyang,Yang, Runjun,Li, Junya,Zhang, Lupei,Liu, Jing,Lu, Chunyan,Lian, Chuanjiang,Li, Zezhong,Zhang, Yong-Hong,Zhang, Liying,Zhao, Zhihui Asian Australasian Association of Animal Productio 2012 Animal Bioscience Vol.25 No.5

        The FAT-1 protein is an n-3 fatty acid desaturase, which can recognize a range of 18- and 20-carbon n-6 substrates and transform n-6 polyunsaturated fatty acids (PUFAs) into n-3 PUFAs while n-3 PUFAs have beneficial effect on human health. Fat1 gene is the coding sequence from Caenorhabditis elegans which might play an important role on lipometabolism. To reveal the function of fat1 gene in bovine fetal fibroblast cells and gain the best cell nuclear donor for transgenic bovines, the codon of fat1 sequence was optimized based on the codon usage frequency preference of bovine muscle protein, and directionally cloned into the eukaryotic expression vector pEF-GFP. After identifying by restrictive enzyme digests with AatII/XbaI and sequencing, the fusion plasmid pEF-GFP-fat1 was identified successfully. The pEF-GFP-fat1 vector was transfected into bovine fetal fibroblast cells mediated by Lipofectamine2000$^{TM}$. The positive bovine fetal fibroblast cells were selected by G418 and detected by RT-PCR. The results showed that a 1,234 bp transcription was amplified by reverse transcription PCR and the positive transgenic fat1 cell line was successfully established. Then the expression level of fat1 gene in positive cells was detected using quantitative PCR, and the catalysis efficiency was detected by gas chromatography. The results demonstrated that the catalysis efficiency of fat1 was significantly high, which can improve the total PUFAs rich in EPA, DHA and DPA. Construction and expression of pEF-GFP-fat1 vector should be helpful for further understanding the mechanism of regulation of fat1 in vitro. It could also be the first step in the production of fat1 transgenic cattle.

      • SCIESCOPUSKCI등재

        Differential Expression of PPARγ, FASN, and ACADM Genes in Various Adipose Tissues and Longissimus dorsi Muscle from Yanbian Yellow Cattle and Yan Yellow Cattle

        Ji, Shuang,Yang, Runjun,Lu, Chunyan,Qiu, Zhengyan,Yan, Changguo,Zhao, Zhihui Asian Australasian Association of Animal Productio 2014 Animal Bioscience Vol.27 No.1

        The objective of this study was to investigate the correlation between cattle breeds and deposit of adipose tissues in different positions and the gene expressions of peroxisome proliferator-activated receptor gamma ($PPAR{\gamma}$), fatty acid synthase (FASN), and Acyl-CoA dehydrogenase (ACADM), which are associated with lipid metabolism and are valuable for understanding the physiology in fat depot and meat quality. Yanbian yellow cattle and Yan yellow cattle reared under the same conditions display different fat proportions in the carcass. To understand this difference, the expression of $PPAR{\gamma}$, FASN, and ACADM in different adipose tissues and longissimus dorsi muscle (LD) in these two breeds were analyzed using the Real-time quantitative polymerase chain reaction method (qRT-PCR). The result showed that $PPAR{\gamma}$ gene expression was significantly higher in adipose tissue than in LD in both breeds. $PPAR{\gamma}$ expression was also higher in abdominal fat, in perirenal fat than in the subcutaneous fat (p<0.05) in Yanbian yellow cattle, and was significantly higher in subcutaneous fat in Yan yellow cattle than that in Yanbian yellow cattle. On the other hand, FASN mRNA expression levels in subcutaneous fat and abdominal fat in Yan yellow cattle were significantly higher than that in Yanbian yellow cattle. Interestingly, ACADM gene shows greater fold changes in LD than in adipose tissues in Yan yellow cattle. Furthermore, the expressions of these three genes in lung, colon, kidney, liver and heart of Yanbian yellow cattle and Yan yellow cattle were also investigated. The results showed that the highest expression levels of $PPAR{\gamma}$ and FASN genes were detected in the lung in both breeds. The expression of ACADM gene in kidney and liver were higher than that in other organs in Yanbian yellow cattle, the comparison was not statistically significant in Yan yellow cattle.

      • KCI등재

        Deposition of Nd-Doped Fe2O3 Nanoparticles on Cenosphere by Hydrothermal Method

        Hui Zhang,Yuanyuan Shi,Jun Xu,Runjun Sun 성균관대학교(자연과학캠퍼스) 성균나노과학기술원 2016 NANO Vol.11 No.3

        A layer of flake-like Fe2O3 particles doped with rare earth Nd3+ ions is homogeneously coated on the surface of cenosphere by using ferric nitrate as the iron source, tartaric acid as the precipitating agent, hexadecyl trimethyl ammonium bromide as the dispersing agent and neodymium nitrate as the dopant via a facile hydrothermal route. The as-prepared cenosphere is characterized by various analytical techniques such as field emission scanning electron microscopy, energy dispersive X-ray spectroscopy, X-ray diffraction, transmission electron microscopy, vibrating sample magnetometry, thermal gravimetric analysis, differential scanning calorimetry, diffuse reflectance spectroscopy, photoluminescence spectroscopy, Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy. The performances of photocatalytic degradation of methylene blue dye are also investigated under ultraviolet and visible light irradiations. The experimental results indicate that the doping concentration of Nd3+ ions is optimized as 0.4% with respect to Fe3+ ions, and the rare earth Nd3+ ions are highly dispersed onto Fe2O3 particle surface. After being doped with Nd3+ ions, the photoactivity of the 0.4% Nd-doped Fe2O3 coated cenosphere is distinctly improved. The magnetic properties are also enhanced to a large extent.

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