A BIDIRECTIONAL ENHANCER CLONING VEHICLE FOR HIGHER-PLANTS

被引:9
作者
OTT, RW
REN, L
CHUA, NH
机构
[1] Laboratory for Plant Molecular Biology, The Rockefeller University, New York, 10021-6399, NY
来源
MOLECULAR AND GENERAL GENETICS | 1990年 / 221卷 / 01期
关键词
Cloning vehicles; Enhancers; Transgenic plants;
D O I
10.1007/BF00280376
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
We have constructed an enhancer cloning vehicle for higher plants, which is based on Agrobacterium tumefaciens T-DNA-mediated transformation of Nicotiana tabacum leaf disk segments. The binary vector plasmid, pROA97, contains two genes in a divergent transcription pattern, which are separated by a unique Bg/II cloning site. Both genes carry abbreviated TATA regions from the cauliflower mosaic virus 35S promoter region. The neomycin phosphotransferase (NPT II) gene is used as a selectable marker in callus tissue cultured cells. Expression from the other gene encoding β-glucuronidase (GUS) can be monitored in regenerated plant tissues by fluorimetric and histochemical assays. Here we describe the transformation properties of the empty vector and control constructs with either the 35S enhancer or the nopaline synthetase promoter region inserted into the unique Bg/II site. A construct carrying the 35 S - 46 to + 8 TATA region is incapable of allowing callus production on selectable media; a construct carrying the 35 S -90 to + 8 TATA region linked to the NPT II gene coding sequence is sufficient to give transformed callus cells and regenerated transgenic plants. There is a high correlation between expression levels of the two genes in this bidirectional system. The vectors were designed so that the enhancer sequences could be rescued from the plant genome by a rapid marker rescue protocol. © 1990 Springer-Verlag.
引用
收藏
页码:121 / 124
页数:4
相关论文
共 17 条
  • [1] PLASMID VEHICLES FOR DIRECT CLONING OF ESCHERICHIA-COLI PROMOTERS
    AN, G
    FRIESEN, JD
    [J]. JOURNAL OF BACTERIOLOGY, 1979, 140 (02) : 400 - 407
  • [2] GENE TAGGING IN PLANTS BY A T-DNA INSERTION MUTAGEN THAT GENERATES APH(3')II-PLANT GENE FUSIONS
    ANDRE, D
    COLAU, D
    SCHELL, J
    VANMONTAGU, M
    HERNALSTEENS, JP
    [J]. MOLECULAR AND GENERAL GENETICS, 1986, 204 (03): : 512 - 518
  • [3] ANALYSIS OF GENE-CONTROL SIGNALS BY DNA-FUSION AND CLONING IN ESCHERICHIA-COLI
    CASADABAN, MJ
    COHEN, SN
    [J]. JOURNAL OF MOLECULAR BIOLOGY, 1980, 138 (02) : 179 - 207
  • [4] PROMOTER-PROBE PLASMID FOR BACILLUS-SUBTILIS
    DONNELLY, CE
    SONENSHEIN, AL
    [J]. JOURNAL OF BACTERIOLOGY, 1984, 157 (03) : 965 - 967
  • [5] DONNELLY CE, 1982, MOL CLONING GENE REG, P63
  • [6] EXPRESSION OF BACTERIAL GENES IN PLANT-CELLS
    FRALEY, RT
    ROGERS, SG
    HORSCH, RB
    SANDERS, PR
    FLICK, JS
    ADAMS, SP
    BITTNER, ML
    BRAND, LA
    FINK, CL
    FRY, JS
    GALLUPPI, GR
    GOLDBERG, SB
    HOFFMANN, NL
    WOO, SC
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES, 1983, 80 (15): : 4803 - 4807
  • [7] GODINI D, 1988, MOL GEN GENET, V211, P507
  • [8] MOUSE EMBRYONIC STEM-CELLS AND REPORTER CONSTRUCTS TO DETECT DEVELOPMENTALLY REGULATED GENES
    GOSSLER, A
    JOYNER, AL
    ROSSANT, J
    SKARNES, WC
    [J]. SCIENCE, 1989, 244 (4903) : 463 - 465
  • [9] ISOLATION OF TOBACCO DNA SEGMENTS WITH PLANT PROMOTER ACTIVITY
    HERMAN, LMF
    VANMONTAGU, MC
    DEPICKER, AG
    [J]. MOLECULAR AND CELLULAR BIOLOGY, 1986, 6 (12) : 4486 - 4492
  • [10] JEFFERSON RA, 1987, EMBO J, V6, P3901