脂肪鉴定实验是高中生物学的经典实验,教材中多用苏丹Ⅲ、Ⅳ对植物、动物脂肪进行染色鉴定,缺少对动物组织中脂肪的鉴定观察。本文利用石蜡切片苏木精伊红染色(HE染色)和冰冻切片油红O(Oil Red O,ORO)染色法,对脂肪肝小鼠肝脏组织进行...脂肪鉴定实验是高中生物学的经典实验,教材中多用苏丹Ⅲ、Ⅳ对植物、动物脂肪进行染色鉴定,缺少对动物组织中脂肪的鉴定观察。本文利用石蜡切片苏木精伊红染色(HE染色)和冰冻切片油红O(Oil Red O,ORO)染色法,对脂肪肝小鼠肝脏组织进行切片观察,与正常肝组织对比呈现出明显差异。增加实验的探索性,引入不同染色方法在脂肪鉴定中的应用,不仅拓展学生的知识眼界,丰富科创活动,也辅助了课堂教学。展开更多
A full-length sequence coding for △^12 fatty acid desaturase gene from peanut（Arachis hypogaea L.）was cloned into the expression vector, pRSETB, to generate recombinant plasmid pRSET/HO-A, which was subsequently tr...A full-length sequence coding for △^12 fatty acid desaturase gene from peanut（Arachis hypogaea L.）was cloned into the expression vector, pRSETB, to generate recombinant plasmid pRSET/HO-A, which was subsequently transformed into expression Escherichia. coli BL21（DE3）pLysS. The △^12 fatty acid desaturase was highly expressed in E. coli BL21（DE3）pLysS in the presence of isopropyl-D-thiogalactopyranoside （IPTG）. The fusion protein was purified and used to form a reaction system in vitro by adding oleic acid as substrate and incubating it at 20℃ for 6 h. Total fatty acids was extracted and methlesterified and then analyzed with gas chromatography. A novel peak corresponding to linoleic acid methyl ester standards was detected with the same retention time. GC-MS （gas chromatogram and gas chromatogram-mass spectrometry） analysis showed that the novel peak was linoleic acid methyl ester. These results exhibited △^12 fatty acid desaturase activity, which could convert oleic acid to linoleic acid specifically.展开更多
文摘脂肪鉴定实验是高中生物学的经典实验,教材中多用苏丹Ⅲ、Ⅳ对植物、动物脂肪进行染色鉴定,缺少对动物组织中脂肪的鉴定观察。本文利用石蜡切片苏木精伊红染色(HE染色)和冰冻切片油红O(Oil Red O,ORO)染色法,对脂肪肝小鼠肝脏组织进行切片观察,与正常肝组织对比呈现出明显差异。增加实验的探索性,引入不同染色方法在脂肪鉴定中的应用,不仅拓展学生的知识眼界,丰富科创活动,也辅助了课堂教学。
基金This work was supported by Chinese National Programs for High Technology Research and Development (No. 2002AA207004).
文摘A full-length sequence coding for △^12 fatty acid desaturase gene from peanut（Arachis hypogaea L.）was cloned into the expression vector, pRSETB, to generate recombinant plasmid pRSET/HO-A, which was subsequently transformed into expression Escherichia. coli BL21（DE3）pLysS. The △^12 fatty acid desaturase was highly expressed in E. coli BL21（DE3）pLysS in the presence of isopropyl-D-thiogalactopyranoside （IPTG）. The fusion protein was purified and used to form a reaction system in vitro by adding oleic acid as substrate and incubating it at 20℃ for 6 h. Total fatty acids was extracted and methlesterified and then analyzed with gas chromatography. A novel peak corresponding to linoleic acid methyl ester standards was detected with the same retention time. GC-MS （gas chromatogram and gas chromatogram-mass spectrometry） analysis showed that the novel peak was linoleic acid methyl ester. These results exhibited △^12 fatty acid desaturase activity, which could convert oleic acid to linoleic acid specifically.
