Cloning,Expression and Biological Effect of Pineapple Cu/Zn-SOD Superoxide Dismutase
碩士 === 國立海洋大學 === 水產生物技術研究所 === 88 === A full-length cDNA clone of 820 bp encoding a putative copper / zinc-superoxide dismutase ( Cu/Zn-SOD ) from pineapple fruit was cloned by PCR approach. Nucleotide sequence analysis of this cDNA clone revealed that it comprised a complete open readin...
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ndltd-TW-088NTOU06130132016-01-29T04:14:30Z http://ndltd.ncl.edu.tw/handle/74811647007849830996 Cloning,Expression and Biological Effect of Pineapple Cu/Zn-SOD Superoxide Dismutase 鳳梨銅鋅超氧歧化之基因選殖、表現與其生物性效應 Fu Ming Tzu 傅明慈 碩士 國立海洋大學 水產生物技術研究所 88 A full-length cDNA clone of 820 bp encoding a putative copper / zinc-superoxide dismutase ( Cu/Zn-SOD ) from pineapple fruit was cloned by PCR approach. Nucleotide sequence analysis of this cDNA clone revealed that it comprised a complete open reading frame coding for 152 amino acid residues. The deduced amino acid sequences showed high similarity ( 62.5 % ~ 81.6 % ) with the sequences of the Cu/Zn-SODs from other plant species. Computer analysis of the residues required for coordinating copper ( His-45, 47,62 and 119 ) and zinc ( His-62, 70,79 and Asp-82 ), as well as the two cysteines (56 and 145) that form a single disulfide bond, were well-conserved among all reported Cu/Zn-SOD sequences in present study. To further characterize the pineapple Cu/Zn-SOD, we subcloned its cDNA into an expression vector, pGEX-4T-1, and transformed into Escherichia coli XL-1blue.The expression of the recombinant Cu/Zn-SOD cDNA was confirmed by enzyme activity staining on a native acrylamide gel. In addition, the biological effect of the pineapple Cu/Zn-SOD was also investigated by transfecting pcDNA3-Cu/Zn-SOD into the microglial BV-2 cells which were treated lipopolysaccharide ( LPS, from the cell wall of gram-negative bacteria ) after transfection.. We found that expressed pineapple Cu/Zn-SOD in BV-2 cells could change the intracellular redox state and decrease the production of NO·, IL-1 and TNF- . These result suggested that the plant Cu/Zn-SOD may be useful in the reduction of the cellular immune reactions. Lin Chi Tsai 林棋財 2000 學位論文 ; thesis 80 zh-TW |
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碩士 === 國立海洋大學 === 水產生物技術研究所 === 88 === A full-length cDNA clone of 820 bp encoding a putative copper / zinc-superoxide dismutase ( Cu/Zn-SOD ) from pineapple fruit was cloned by PCR approach. Nucleotide sequence analysis of this cDNA clone revealed that it comprised a complete open reading frame coding for 152 amino acid residues. The deduced amino acid sequences showed high similarity ( 62.5 % ~ 81.6 % ) with the sequences of the Cu/Zn-SODs from other plant species. Computer analysis of the residues required for coordinating copper ( His-45, 47,62 and 119 ) and zinc ( His-62, 70,79 and Asp-82 ), as well as the two cysteines (56 and 145) that form a single disulfide bond, were well-conserved among all reported Cu/Zn-SOD sequences in present study. To further characterize the pineapple Cu/Zn-SOD, we subcloned its cDNA into an expression vector, pGEX-4T-1, and transformed into Escherichia coli XL-1blue.The expression of the recombinant Cu/Zn-SOD cDNA was confirmed by enzyme activity staining on a native acrylamide gel. In addition, the biological effect of the pineapple Cu/Zn-SOD was also investigated by transfecting pcDNA3-Cu/Zn-SOD into the microglial BV-2 cells which were treated lipopolysaccharide ( LPS, from the cell wall of gram-negative bacteria ) after transfection.. We found that expressed pineapple Cu/Zn-SOD in BV-2 cells could change the intracellular redox state and decrease the production of NO·, IL-1 and TNF- . These result suggested that the plant Cu/Zn-SOD may be useful in the reduction of the cellular immune reactions.
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Lin Chi Tsai |
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Lin Chi Tsai Fu Ming Tzu 傅明慈 |
author |
Fu Ming Tzu 傅明慈 |
spellingShingle |
Fu Ming Tzu 傅明慈 Cloning,Expression and Biological Effect of Pineapple Cu/Zn-SOD Superoxide Dismutase |
author_sort |
Fu Ming Tzu |
title |
Cloning,Expression and Biological Effect of Pineapple Cu/Zn-SOD Superoxide Dismutase |
title_short |
Cloning,Expression and Biological Effect of Pineapple Cu/Zn-SOD Superoxide Dismutase |
title_full |
Cloning,Expression and Biological Effect of Pineapple Cu/Zn-SOD Superoxide Dismutase |
title_fullStr |
Cloning,Expression and Biological Effect of Pineapple Cu/Zn-SOD Superoxide Dismutase |
title_full_unstemmed |
Cloning,Expression and Biological Effect of Pineapple Cu/Zn-SOD Superoxide Dismutase |
title_sort |
cloning,expression and biological effect of pineapple cu/zn-sod superoxide dismutase |
publishDate |
2000 |
url |
http://ndltd.ncl.edu.tw/handle/74811647007849830996 |
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