ORGAN-SPECIFIC MODULATION OF GENE-EXPRESSION IN TRANSGENIC PLANTS USING ANTISENE RNA

被引:51
作者
CANNON, M
PLATZ, J
OLEARY, M
SOOKDEO, C
CANNON, F
机构
[1] Bio Technica International, Inc., Cambridge, 02140, MA
[2] Plant Pathology Dept., University of Massachusetts, Amherst, 01003, MA
[3] Dept. of Molecular Biology, Massachusetts General Hospital, Boston, 02114, MA
关键词
antisense; gene expression; plants; regulation; RNA; transformation;
D O I
10.1007/BF00017722
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 shown leaf-specific inhibition GUS gene expression in transgenic Nicotiana plants using an antisense RNA with a 41-base homology spanning the translation start codon of the gene. GUS was expressed from the nominally constitutive 35S promoter and the antisense RNA was expressed from the light-regulated ca/b promoter of Arabidopsis thaliana. A range of GUS inhibition from 0 to 100% was obtained by screening a small population of transgenic plants and the specific levels of inhibition observed were stably inherited in two generations. An antiGUS 'gene' dosage effect was observed in plants which were homozygous for antiGUS. RNA detection results suggest that duplex formation with the 41 base pair antiGUS RNA destabilized the GUS mRNA and that an excess of antisense. RNA was not required. Our results demonstrate the potential of antisense RNA as a strategy for obtaining plant mutants, especially 'down mutations' in essential genes where only a short 5′ sequence of the mRNA is required. They also suggest that the 'position effect' on gene expression could be used in conjunction with an antisense RNA strategy to provide a versatile approach for crop improvement. © 1990 Kluwer Academic Publishers.
引用
收藏
页码:39 / 47
页数:9
相关论文
共 31 条
[1]  
Ausubel FM., 1995, MOL REPROD DEV, V3rd edn, DOI DOI 10.1002/MRD.1080010210
[2]   BINARY AGROBACTERIUM VECTORS FOR PLANT TRANSFORMATION [J].
BEVAN, M .
NUCLEIC ACIDS RESEARCH, 1984, 12 (22) :8711-8721
[3]   A COMPLEMENTATION ANALYSIS OF RESTRICTION AND MODIFICATION OF DNA IN ESCHERICHIA COLI [J].
BOYER, HW ;
ROULLAND.D .
JOURNAL OF MOLECULAR BIOLOGY, 1969, 41 (03) :459-&
[4]   THE MOB AND ORIT MOBILIZATION FUNCTIONS OF A BACTERIAL PLASMID PROMOTE ITS TRANSFER TO PLANTS [J].
BUCHANANWOLLASTON, V ;
PASSIATORE, JE ;
CANNON, F .
NATURE, 1987, 328 (6126) :172-175
[5]  
COLMAN J, 1985, NATURE, V315, P601
[6]  
CROWLEY TE, 1985, CELL, V43, P33
[7]   A STABLE BIFUNCTIONAL ANTISENSE TRANSCRIPT INHIBITING GENE-EXPRESSION IN TRANSGENIC PLANTS [J].
DELAUNEY, AJ ;
TABAEIZADEH, Z ;
VERMA, DPS .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1988, 85 (12) :4300-4304
[8]   IDENTIFICATION OF SEQUENCES INVOLVED IN THE POLYADENYLATION OF HIGHER-PLANT NUCLEAR TRANSCRIPTS USING AGROBACTERIUM T-DNA GENES AS MODELS [J].
DHAESE, P ;
DEGREVE, H ;
GIELEN, J ;
SEURINCK, J ;
VANMONTAGU, M ;
SCHELL, J .
EMBO JOURNAL, 1983, 2 (03) :419-426
[9]   BROAD HOST RANGE DNA CLONING SYSTEM FOR GRAM-NEGATIVE BACTERIA - CONSTRUCTION OF A GENE BANK OF RHIZOBIUM-MELILOTI [J].
DITTA, G ;
STANFIELD, S ;
CORBIN, D ;
HELINSKI, DR .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES, 1980, 77 (12) :7347-7351
[10]  
DUNSMUIR P, 1988, PLANT MOL BIOL MAN C, V1, P1