Isolation of proteins and nucleic acids by electrophoresis on disposable gel columns

被引:1
作者
Costa, MA
Duro, G
Porcasi, R
Colombo, P
Locorotondo, G
Geraci, D
Izzo, V
机构
[1] Istituto di Biologia dello Sviluppo, CNR, Palermo
[2] Istituto di Biologia dello Sviluppo, CNR, 90123 Palermo
关键词
electrophoresis in disposable gel columns; DNA purification; protein purification; disposable gel column;
D O I
10.1002/elps.1150170427
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
A simple and cheap one-step method to isolate proteins or nucleic acids by electrophoresis in disposable gel columns is reported. A disposable syringe was modified to host a gel column and an elution chamber. Starting from a crude extract of E. coli, the laboratory-made devise allowed the isolation of the maltose binding protein (MBP) fused to a recombinant allergenic molecule with a molecular mass of 58 kDa, from a mixture of several proteins. Also, plasmid DNA could be isolated from a mixture containing chromosomal DNA and RNA, avoiding the use of organic solvents. Electrophoresis was performed at 150 V, 35 degrees C, pH 8.0 and 8.3 for protein and DNA, respectively. The protein or the DNA obtained showed a yield of 80% and a purity grade of 90%, as estimated by densitometry.
引用
收藏
页码:781 / 783
页数:3
相关论文
共 9 条
  • [1] [Anonymous], 1988, Antibodies: A Laboratory Manual
  • [2] ELECTROPHORETIC PARTICLE SIEVING IN POLYACRYLAMIDE GELS AS APPLIED TO RIBOSOMES
    HJERTEN, S
    JERSTEDT, S
    TISELIUS, A
    [J]. ANALYTICAL BIOCHEMISTRY, 1965, 11 (02) : 211 - &
  • [3] 2 METHODS FOR ELECTROPHORETIC ELUTION OF PROTEINS FROM POLYACRYLAMIDE GELS
    KARSNAS, P
    ROOS, P
    [J]. ANALYTICAL BIOCHEMISTRY, 1977, 77 (01) : 168 - 175
  • [4] OLECHNO JD, 1991, AM LAB MAR, P59
  • [5] Sambrook J., 2002, MOL CLONING LAB MANU
  • [6] ELECTROFRACTIONATION - A TECHNIQUE FOR DETECTING AND RECOVERING BIOMOLECULES
    SHAIN, DH
    YOO, JY
    SLAUGHTER, RG
    HAYES, SE
    JI, TH
    [J]. ANALYTICAL BIOCHEMISTRY, 1992, 200 (01) : 47 - 51
  • [7] SHEER DG, 1990, BIOTECHNIQUES, V9, P486
  • [8] IMMUNOAFFINITY CHROMATOGRAPHY USING ELECTROELUTION
    WILLIAMSON, KC
    DUFFY, PE
    KASLOW, DC
    [J]. ANALYTICAL BIOCHEMISTRY, 1992, 206 (02) : 359 - 362
  • [9] ELECTROELUTION AND PURIFICATION OF CHLOROPLAST DNA SUITABLE FOR RESTRICTION ANALYSES
    WONG, VWT
    FONES, AML
    LOWREY, TK
    [J]. ANALYTICAL BIOCHEMISTRY, 1991, 194 (02) : 378 - 380