Maize genes for controlling plant growth and organ size and their use in improving crop plants
A plant, corn technology, applied in the field of molecular biology, can solve the problem of undetected
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Embodiment 1
[0234] Example 1. Isolation of ARGOS sequences
[0235] Related ARGOS genes were found using conventional methods for identifying all members of a gene family. The protein sequences of all known members of the various groups of this gene family were prepared. This data includes sequences from other species. These species were searched in proprietary maize sequence datasets and non-redundant sets of overlapping results were identified. In a stepwise manner, the nucleotide sequence of any gene of interest is obtained, the database is searched and a non-redundant set of all overlapping results is retrieved. These sets of protein results are then compared to the nucleotide results. If the gene family is complete, all protein results are included in the nucleotide results. The ARGOS gene family consists of 3 Arabidopsis genes, 8 rice genes, 9 maize genes, 9 sorghum genes and 5 soybean genes. For a dendrogram illustration of the interrelationships of proteins encoded by the...
Embodiment 2
[0236] Example 2. ARGOS sequence analysis
[0237] The ZmARGOS polypeptide of the present invention has common characteristics with ARGOS genes of various plant species. The relationship of the genes in multiple plant species is expressed in the form of comparison, as shown in Figure 2. image 3 Contains ZmARGOS 1, 2, 3 and AtARGOS1 (SEQ ID NO: 2, 4, 6 and 26). The protein encoded by the ARGOS gene has a highly conserved proline-rich region near the C-terminus. The N-terminus is more variable. The protein is relatively short, containing an average of 110 amino acids.
Embodiment 3
[0238] Example 3. Transformation and regeneration of transgenic plants
[0239] Bombardment with a plasmid containing the ZmARGOS sequence operably linked to the drought-inducible promoter RAB17 promoter (Vilardell et al. (1990) Plant Mol Biol 14:423-432) and the selectable marker gene PAT (which confers resistance to the herbicide Bialaphos) Immature maize embryos from greenhouse donor plants. Alternatively the selectable marker gene can be provided on a separate plasmid. Transform according to the following steps. The medium recipe is shown below.
[0240] Preparation of target tissue:
[0241] Ears were dehulled and surface sterilized with 30% Cloros bleach plus 0.5% Micro detergent for 20 minutes and rinsed twice with sterile water. Immature embryos were minced and placed axis-side down (scutellum side up), 25 embryos per plate, placed in 560Y medium for 4 hours and then arrayed in a 2.5-cm target area for bombardment .
[0242] DNA preparation
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