Parameters influencing ectopic gene expression in Aplysia neurons

被引:57
作者
Kaang, BK
机构
[1] Molecular Neurobiology Laboratory, Inst. for Molec. Biol. and Genetics, Seoul National University, Seoul, 151-742, San 56-1, Silim-dong, Kwanak-gu
关键词
Aplysia neurons; ganglion culture; gene transfer; DNA microinjection; AP-1 enhancer element; transcription factor; beta-galactosidase; firefly luciferase;
D O I
10.1016/S0304-3940(96)13279-1
中图分类号
Q189 [神经科学];
学科分类号
071006 ;
摘要
DNA microinjection for expressing exogenous genes in Aplysia neurons has been very useful for analyzing not only functions of encoded proteins, but also regulations of gene expression in the nervous system. Some of the factors that affect expression of foreign genes microinjected into Aplysia neurons are described. The effect of the DNA form (supercoiled or linear) and promoter modification in the expression vectors are analyzed as well as buffer composition and DNA concentration in the microinjection solution. The time course of reporter gene expression was also monitored. Reporter gene expression was first detected as early as 1 h and maintained at a high level even until 7 days after microinjection. The presence of AP-1 enhancer in the promoter region of the expression vectors was essential in driving a high-level constitutive expression of reporter genes. Particularly, a pNEX derivative containing eight copies of AP-1 enhancer drove constitutive overexpression more effectively than ones harboring either four or 12 copies of AP-1 enhancer. We also found that a prokaryotic promoter/operator from E. coli lacZ gene placed upstream from an eukaryotic enhancer/promoter repressed the expression of the downstream reporter gene in Aplysia neurons. (C) 1996 Elsevier Science Ireland Ltd.
引用
收藏
页码:29 / 32
页数:4
相关论文
共 14 条
  • [1] [Anonymous], [No title captured]
  • [2] APLYSIA-CALIFORNICA NEURONS EXPRESS MICROINJECTED NEUROPEPTIDE GENES
    DESGROSEILLERS, L
    COWAN, D
    MILES, M
    SWEET, A
    SCHELLER, RH
    [J]. MOLECULAR AND CELLULAR BIOLOGY, 1987, 7 (08) : 2762 - 2771
  • [3] A DEFECTIVE HSV-1 VECTOR EXPRESSES ESCHERICHIA-COLI BETA-GALACTOSIDASE IN CULTURED PERIPHERAL NEURONS
    GELLER, AI
    BREAKEFIELD, XO
    [J]. SCIENCE, 1988, 241 (4873) : 1667 - 1669
  • [4] THE ROUS-SARCOMA VIRUS LONG TERMINAL REPEAT IS A STRONG PROMOTER WHEN INTRODUCED INTO A VARIETY OF EUKARYOTIC CELLS BY DNA-MEDIATED TRANSFECTION
    GORMAN, CM
    MERLINO, GT
    WILLINGHAM, MC
    PASTAN, I
    HOWARD, BH
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES, 1982, 79 (22): : 6777 - 6781
  • [5] Voltage-dependent modulation of N-type calcium channels by G-protein beta gamma subunits
    Ikeda, SR
    [J]. NATURE, 1996, 380 (6571) : 255 - 258
  • [6] ACTIVATION OF CAMP-RESPONSIVE GENES BY STIMULI THAT PRODUCE LONG-TERM FACILITATION IN APLYSIA SENSORY NEURONS
    KAANG, BK
    KANDEL, ER
    GRANT, SGN
    [J]. NEURON, 1993, 10 (03) : 427 - 435
  • [7] Kaang BK, 1996, MOL CELLS, V6, P285
  • [8] OVEREXPRESSION OF AN APLYSIA SHAKER K+ CHANNEL GENE MODIFIES THE ELECTRICAL-PROPERTIES AND SYNAPTIC EFFICACY OF IDENTIFIED APLYSIA NEURONS
    KAANG, BK
    PFAFFINGER, PJ
    GRANT, SGN
    KANDEL, ER
    FURUKAWA, Y
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1992, 89 (03) : 1133 - 1137
  • [9] MOLECULAR-BIOLOGY OF LEARNING - MODULATION OF TRANSMITTER RELEASE
    KANDEL, ER
    SCHWARTZ, JH
    [J]. SCIENCE, 1982, 218 (4571) : 433 - 443
  • [10] AN ADENOVIRUS VECTOR FOR GENE-TRANSFER INTO NEURONS AND GLIA IN THE BRAIN
    LASALLE, GL
    ROBERT, JJ
    BERRARD, S
    RIDOUX, V
    STRATFORDPERRICAUDET, LD
    PERRICAUDET, M
    MALLET, J
    [J]. SCIENCE, 1993, 259 (5097) : 988 - 990