Abstract
Previous studies have shown that a chimeric streptomycin phosphotransferase (SPT) gene can function as a dominant marker for plant cell transformation. The SPT marker previously described by Jones and co-workers has a limited value since it conferred a useful level of resistance only to a fraction (10%) of Nicotiana plumbaginifolia transgenic lines. Expression of resistance was species specific: no such resistant transformants were found in N. tabacum. In this paper we describe an improved SPT construct that utilizes a mutant Tn5 SPT gene. The mutant gene, SPT*, encodes a protein with a two amino acid deletion close to its COOH-terminus. In N. tabacum cell culture the efficiency of transformation with the improved streptomycin resistance marker was comparable to kanamycin resistance. When the chimeric SPT* gene was introduced linked to a kanamycin resistance gene, streptomycin resistance was expressed in most of the transgenic N. tabacum lines.
MeSH Terms
Chimera
DNA, Recombinant
Gene Expression Regulation
Genetic Markers
Genetic Vectors
Kanamycin Resistance/genetics
Mutation
Phosphotransferases/genetics
Phosphotransferases (Alcohol Group Acceptor)
Plants, Toxic
Plasmids
Tobacco/genetics
Transformation, Genetic
Chemicals
DNA, Recombinant
Genetic Markers
Phosphotransferases
Phosphotransferases (Alcohol Group Acceptor)
streptomycin 3''-kinase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Maliga P
Advanced Genetic Sciences Inc., Oakland, CA 94608.
Svab Z
Harper E C
Jones J D
References (9)
9 references, click to expand
-
Tn5 carries a streptomycin resistance determinant downstream from the kanamycin resistance gene.
Mol Gen Genet. 1983;191(2):288-94
PMID: 6312272
-
Integration and organization of Ti plasmid sequences in crown gall tumors.
Cell. 1980 Mar;19(3):729-39
PMID: 7363328
-
Photoregulated expression of a pea rbcS gene in leaves of transgenic plants.
EMBO J. 1985 Dec 1;4(12):3063-8
PMID: 16453648
-
Expression of Tn5-encoded streptomycin resistance in E. coli.
Mol Gen Genet. 1986 Sep;204(3):404-9
PMID: 3020378
-
Simple binary vectors for DNA transfer to plant cells.
Plant Mol Biol. 1985 May;5(3):149-54
PMID: 24306650
-
High level expression of introduced chimaeric genes in regenerated transformed plants.
EMBO J. 1985 Oct;4(10):2411-8
PMID: 15929216
-
The ocs element: a 16 base pair palindrome essential for activity of the octopine synthase enhancer.
EMBO J. 1987 Nov;6(11):3203-8
PMID: 16453801
-
Isolation of a dual plant promoter fragment from the Ti plasmid of Agrobacterium tumefaciens.
EMBO J. 1984 Dec 1;3(12):2723-30
PMID: 16453574
-
The complete nucleotide sequence of the TL-DNA of the Agrobacterium tumefaciens plasmid pTiAch5.
EMBO J. 1984 Apr;3(4):835-46
PMID: 6327292