[1]程光宇,王 峰,陶明煊,等.聚丙烯酰胺梯度凝胶电泳和均一凝胶电泳在分离及鉴定植物SOD同工酶中的比较研究[J].南京师大学报(自然科学版),2008,31(04):97-103.
　Cheng Guangyu,Wang Feng,Tao Minxuan,et al.Comparison of Gradient Polyacrylamide and Polyacrylamide Gel Electrophoresis in the Isolation and Identification of Plant SOD Isozyme[J].Journal of Nanjing Normal University(Natural Science Edition),2008,31(04):97-103.
点击复制

聚丙烯酰胺梯度凝胶电泳和均一凝胶电泳在分离及鉴定植物SOD同工酶中的比较研究()
分享到：

《南京师大学报（自然科学版）》[ISSN:1001-4616/CN:32-1239/N]

卷:
第31卷
期数:
2008年04期
页码:
97-103
栏目:
生命科学
出版日期:
2008-12-30

文章信息/Info

Title:
Comparison of Gradient Polyacrylamide and Polyacrylamide Gel Electrophoresis in the Isolation and Identification of Plant SOD Isozyme
作者:
程光宇1;  王  峰2;  陶明煊2;  陆长梅1;  吴国荣1;  常福辰1
( 1. 南京师范大学生命科学学院, 江苏南京210097)
( 2. 南京师范大学金陵女子学院, 江苏南京210097)
Author(s):
Cheng Guangyu1; Wang Feng2; Tao Minxuan2; Lu Changmei1; Wu Gongrong1; Chang Fuchen1
1.School of Life Science,Nanjing Normal University,Nanjing 210097,China
2. Jin l ing C ollege, N an jing Norm alUn iversity, Nan jing 210097, Ch ina
关键词:
聚丙烯酰胺梯度凝胶电泳;  聚丙烯酰胺凝胶电泳;  SOD;  分离;  鉴定
Keywords:
GPGE;  PAGE;  SOD;  Iso lation;  Identifica tion
分类号:
Q946
摘要:
比较了聚丙烯酰胺梯度凝胶电泳( GPGE )和均一凝胶电泳( PAGE )在分离和鉴定SOD同工酶方面的应用. 在GPGE中根据Rf 值大小可将SOD 谱带分为3个区: A区Rf 值最小, 对SDS敏感, 为M n-SOD; C 区Rf 值最大, 对SDS不敏感, 为CuZn-SOD; B区谱带Rf 值位于两者之间, 对SDS敏感, 为Fe-SOD. 根据GPGE 图谱上谱带Rf 值大小和对SDS敏感性来判断SOD类型结果与用抑制剂判断结果一致, 它可作为一种SOD类型初步鉴定的简便可行的方法, 相同植物材料经PAGE分离后不具备这些特性. 在相同牛血SOD浓度下, GPGE检测SOD灵敏度可达10 -15 m o ,l PAGE为10 -13 m ol;在1.56× 10 -12 mol SOD时, 在GPGE中可分辨出4条带, PAGE 仅为2条. 枸杞Fe-SOD 在7. 5% 胶中Rf 值最大, 在10%胶中与CuZn-SOD重叠, 香橼Fe-SOD、Mn-SOD 和朱桔M n-SOD、 CuZn-SOD在10% PAGE中相互重叠, 在GPGE中它们都有规律地完全分离, 可根据Rf 值大小鉴别SOD类型. 通过2次电泳阐明了GPGE 分离和鉴定SOD 的可靠性, 它能有效地分离和鉴定在PAGE 上重叠的3种SOD类型.
Abstract:
Th is paper compared the app lication of grad ient po ly acry lam ide ge l e lectrophresis( GPGE) and po lyacrylam ide ge l e lectrophoresis( PAGE) in iso lation and identifica tion o f plant SOD iso zym e. The resu lts show ed that SOD bands on GPGE w ere div ided into three reg ion of A, B, C accord ing to itsRf. TheRf o fA reg ion SOD band was m inim um, it w as sensitive to 1% SDS and w asM n-SOD. The Rf o f C reg ion SOD band w as the biggest, it was no t sensitive to 1% SDS and w as CuZn-SOD. The Rf of B reg ion SOD band was betw een A and C, it was sensitiv e to 1% SDS and was Fe-SOD. The three SOD iso zym es w ere d istinguished by its Rf and sensitiv ity to SDS on polyacry lam ide g radient ge,l th is resu lt w as about the sam e as the resu lt of judgem en t using inh ibitor. The sam e plant sam plew as iso lated by PAGE d id not have the charac teristic. 10 - 13 m o l and 10 - 15 m ol bov ine SOD w as detected in PAGE and GPGE, respective ly. Four and tw o SOD activ ity bands w ere detected in GPGE and PAGE at 1.56 ×10 - 12 m o l bov ine SOD, respectiv ely. TheRf of Fe-SOD from Lycium chinense w as the bigg est in 7. 5% PAGE and it ove rlapped w ith CuZn-SOD in 10% PAGE. The FeSOD and M n-SOD o f cirus w ilsonii, Mn-SOD and CuZn-SOD of C itrus ery throsa w ere ove rlapped each o ther in 10% PAGE, but three types o f SOD w ere com plete ly iso la ted in 4% ~ 35% GPGE. The re liab ility o f iso lating and identify SOD using GPGE w as expounded by tw o tim es g el concentration grad ient electrophores is TGCGE, the Rf and the num be r of SOD bands in GPGE w ere com patible w ith TGCGE, and it can com plete ly iso late and identify the overlapped three SOD types on PAGE.

参考文献/References:

[ 1] In-Kwon K im, Y ang-In Y im, Young M in-K im, et a.l Cb iX-hom o logous pro te in ( Cb iXhp), a metal-bind ing pro tein, from streptom yces seou lensis is invo lved in expression o f nickel-conta in ing superox ide dismutase[ J] . FEM SM icrob io logy Letters,2003, 228 ( 1): 21-26.
[ 2] Yao C L, W ang A L, W ang Z Y, et a.l Purification and partial character ization o f Cu, Zn superox ide dism utase from haem olymph of orienta l r iver prawn m acrobrachium n ipponense[ J]. Aquacu lture, 2007( 270): 559-565.
[ 3] 罗广华, 王爱国. 植物SOD的凝胶电泳及活性显示[ J] . 植物生理学通讯, 1983( 6): 44-45.
[ 4] 程光宇, 魏锦城, 吴国荣, 等. 枸杞果实铁型超氧物歧化酶的纯化及性质[ J]. 植物生理学报, 1994, 20( 2): 128-136.
[ 5] 王亦根, 程光宇, 吴国荣, 等. 牛血红细胞铜锌超氧物歧化酶的纯化及性质[ J]. 南京师大学报: 自然科学版, 1994, 17( 2): 108-112.
[ 6] 程光宇, 高兴, 洑志清, 等. 花生种子铜锌超氧物歧化酶的纯化及理化性质研究[ J]. 食品工业科技, 2007, 28( 2): 82-85.
[ 7] 程光宇, 魏锦城, 吴国荣. SOD的聚丙稀酰胺梯度凝胶电泳分离和酶活性显示[ J]. 植物生理学通讯, 1994, 30( 4):248.
[ 8] 张龙翔. 生化实验方法和技术[M ]. 北京: 人民教育出版社, 1982: 119-123.
[ 9] 程光宇, 吴国荣, 储慧君. 朱桔M n-SOD的纯化、鉴定及浓度梯度胶电泳在其中的应用[ J]. 植物研究, 2004, 24( 2):240-244.
[ 10] Br idges SM, SalinM L. Distr ibution of iron-containing supe rox ide dismutase in v ascu la r plants[ J]. Plant Phy sio ,l 1981
( 68): 275-278.
[ 11] 方允中, 郑荣梁. 自由基生物学的理论与应用[M ]. 北京: 科学出版社, 2002: 162-178.
[ 12] 赵厚安, 方允中. 锰— 超氧化物歧化酶活力测定的五种方法比较研究[ J] . 生物化学杂志, 1988, 4( 1): 29-35.
[ 13] 罗广华, 王爱国, 付爱根. 鉴别超氧化物歧化酶类型的定位染色法[ J]. 生物化学与生物物理进展, 1996, 23( 4):356-359.
[ 14] 吴国荣, 程光宇, 陆长梅, 等. Fe型超氧物歧化酶在高等维管植物中分布的研究[ J]. 中国野生植物资源, 2001, 19( 3) : 14-16.
[ 15] Kw iatow sk i J, Safianow skaA, K an iug a Z. Iso lation and character iza tion o f an iron-conta in ing supe rox ide d ism utase from to mato leav es, Lycoersicon sca lentun [ J]. Eur J B iochem, 1985, 146: 459-466.
[ 16] Sa linM L, B ridges SM. Iso la tion and charac terization o f an iron-conta in ing superox ide d ism utase from w ater lily, Nuphar luteum [ J]. Plant Physio ,l 1982( 69): 161-165.
[ 17] Sa linM L, Br idges SM. Isolation and charac teriza tion o f an iron-conta ining superox ide d ism utase from a eucaryote, Brassica cam pestr is [ J]. A rch Biochem B iophys, 1980, 201( 2): 369-374.
[ 18] Sa linM L, DukeM V. Purifica tion and character ization o f an iron-conta in ing superox ide dism utase from a eucaryote, G ink-gob iloba[ J]. Arch B iochem Biophys, 1985, 243( 1) : 305-314.
[ 19] Chen H ua iyang, H u Ronggu,i W ang Baozhong, et a.l S tructural studies o f an eukaryo tic cam bia listic superox ide d ism utase purified from them ature seeds of camphor tree[ J] . Arch B iochem B iophys, 2002, 404( 2): 218-226.
[ 20] 罗广华, 王爱国. 植物SOD 同工酶活性显示的某些干扰[ J]. 植物生理学通讯, 1993, 29( 2): 119-122.

备注/Memo

备注/Memo:
基金项目: 江苏省自然科学基金( 1998SWXOSZ001 )资助项目.
通讯联系人: 程光宇, 高级实验师, 研究方向: 生物活性物质及保健功能因子. E-mail:mxwcgy@ yahoo. com. cn
更新日期/Last Update: 2013-05-05