共查询到7条相似文献,搜索用时 0 毫秒
1.
Coutu C Brandle J Brown D Brown K Miki B Simmonds J Hegedus DD 《Transgenic research》2007,16(6):771-781
We present a series of 14 binary vectors suitable for Agrobacterium-mediated transformation of dicotyledonous plants and adaptable for biolistic transformation of monocotyledonous plants. The
vector size has been minimized by eliminating all non-essential elements from the vector backbone and T-DNA regions while
maintaining the ability to replicate independently. The smallest of the vector series is 6.3 kb and possesses an extensive
multiple cloning site with 21 unique restriction endonuclease sites that are compatible with common cloning, protein expression,
yeast two-hybrid and other binary vectors. The T-DNA region was engineered using a synthetic designer oligonucleotide resulting
in an entirely modular system whereby any vector element can be independently exchanged. The high copy number ColE1 origin
of replication has been included to enhance plasmid yield in Escherichia coli. FRT recombination sites flank the selectable marker cassette regions and allow for in planta excision by FLP recombinase. The pORE series consists of three basic types; an ‘open’ set for general plant transformation, a ‘reporter’ set for promoter analysis and an ‘expression’ set for constitutive expression of transgenes. The sets comprise various combinations of promoters (P
HPL, P
ENTCUP2 and P
TAPADH), selectable markers (nptII and pat) and reporter genes (gusA and smgfp). 相似文献
2.
Paula M. Olhoft Libby M. Bernal Leslie B. Grist D. Steven Hill S. Luke Mankin Yuwei Shen Mary Kalogerakis Hunt Wiley Effie Toren Hee-Sook Song Helke Hillebrand Todd Jones 《In vitro cellular & developmental biology. Plant》2007,43(6):536-549
A novel Agrobacterium rhizogenes-mediated transformation method using a primary-node explant from Dairyland cultivar 93061 was developed for soybean using
the disarmed Agrobacterium strain SHA17. Transformed plants regenerated from explants inoculated with SHA17 were fertile and phenotypically normal.
In a comparative experiment, regeneration frequencies were not significantly different between explants inoculated with A. rhizogenes strain SHA17 and Agrobacterium tumefaciens strain AGL1; however, a 3.5-fold increase in transformation efficiency [(number of Southern or TaqMan-positive independent
events/total number of explants inoculated) × 100] was found for explants cocultured with SHA17 compared to AGL1 (6.6 and
1.64%, respectively). Southern analysis of 48 T0 plants suggested that 37.5, 23, and 39.6% of the T0 plants contained 1, 2, and 3 or more T-DNA fragments integrated into the genome, respectively. Additionally, T1 progeny analysis of 8 independent events resulted in typical Mendelian inheritance of T-DNA genes. Of seven T0 plants that had two or more T-DNA fragments, six contained multiple loci segregating in T1 progenies. Further analysis of four lines confirmed the presence of PAT, GUS, and/or DsRED2 proteins in transgenic plants
that were encoded on the T-DNA into the T2 generation. 相似文献
3.
We assessed the effect of four different virulence (vir) gene combinations on plant transformation efficiency and transgene behaviour in rice using the pGreen/pSoup dual binary vector system. Transformation experiments were conducted using a pGreen vector containing the bar and gusA expression units with, or without, the virG542, virGN54D, virGwt or the virG/B/C genes added to the backbone. Additonal vir gene(s) significantly altered plant transformation efficiency and the integration of vector backbone sequences. However, no differences in transgene copy number, percentage of expressing lines and expression levels could be detected. Addition of virGwt was the most beneficial, doubling the overall performance of the pGreen/pSoup vector system based on transformation frequency, absence of backbone sequence integration and expression of unselected transgenes. In 39 of the plant lines, the additional vir genes were integrated into the rice genome. The contribution of super dual binary pGreen/pSoup vectors to the development of efficient rice transformation systems and to the production of plants free of selectable marker genes are discussed. 相似文献
4.
GAs are plant hormones that play fundamental roles in plant growth and development. GA2ox, GA3ox, and GA20ox are three key enzymes in GA biosynthesis. These enzymes belong to the 2OG-Fe (II) oxygenase superfamily and are independently encoded by different gene families. To date, genome-wide comparative analyses of GA oxidases in plant species have not been thoroughly carried out. In the present work, 61 GA oxidase family genes from rice (Oryza sativa), Arabidopsis, and soybean (Glycine max) were identified and a full study of these genes including phylogenetic tree construction, gene structure, gene family expansion and analysis of functional motifs was performed. Based on phylogeny, most of the GA oxidases were divided into four subgroups that reflected functional classifications. Intron/intron average length of GA oxidase genes in rice analysis revealed that GA oxidase genes in rice experienced substantial evolutionary divergence. Segmental duplication events were mainly found in soybean genome. However, in rice and Arabidopsis, no single expansion pattern exhibited dominance, indicating that GA oxidase genes from these species might have been subjected to a more complex evolutionary mechanism. In addition, special functional motifs were discovered in GA20ox, GA3ox, and GA2ox, which suggested that different functional motifs are associated with differences in protein function. Taken together our results suggest that GA oxidase family genes have undergone divergent evolutionary routes, especially at the monocot-dicot split, with dynamic evolution occurring in Arabidopsis thaliana and soybean. 相似文献
5.
Brian A. McGaw Roger Horgan Jim K. Heald George J. Wullems Rob A. Schilperoort 《Planta》1988,176(2):230-234
The levels of the major cytokinins, zeatin, zeatin riboside, zeatin riboside-5-monophosphate and zeatin-7-glucoside were measured in tobacco (Nicotiana tabacum L.) crown-gall tissues carrying insertion and deletion mutations in the T-DNA. Measurements were made by combined gas chromatography-mass spectrometry using selected ion monitoring with 15N- and 2H-labelled internal standards. The results demonstrate that, relative to wild-type tumour tissue, cytokinin levels are considerably elevated in tissues lacking functional T-DNA auxin-biosynthetic genes. From a detailed analysis of the major cytokinin metabolites it is concluded that a reduction in the extent of cytokinin degradation via N6-side-chain cleavage is an important factor leading to increased cytokinin levels in these tissues.Abbreviations IAA
indole-3-acetic acid
- SIM
selected ion monitoring
- Z
zeatin
- [7G]Z
zeatin-7-glucoside
- [9R]Z
zeatin-9-riboside
- [9R-5P]Z
zeatin riboside-5-monophosphate 相似文献
6.