A study on optimization of pat gene expression cassette for maize transformation
| Autor: | Qiudeng Que, Samson Nalapalli, Daolong Wang, Tara Liebler, Tim Strebe, Lee Richbourg, Elumalai Sivamani, David Bradley, Anna Prairie |
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| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
animal structures Agrobacterium Gene Expression Genetically modified crops Zea mays 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Transformation Genetic Acetyltransferases Gene Expression Regulation Plant Genetics Promoter Regions Genetic Molecular Biology Selectable marker biology Herbicides Aminobutyrates General Medicine biology.organism_classification Plants Genetically Modified Transformation (genetics) 030104 developmental biology Glufosinate chemistry Genetic marker 030220 oncology & carcinogenesis cardiovascular system Phosphinothricin acetyltransferase Transformation efficiency |
| Zdroj: | Molecular biology reports. 46(3) |
| ISSN: | 1573-4978 |
| Popis: | Phosphinothricin acetyltransferase gene (pat) is an important selectable marker and also a key herbicide trait gene in several commercial products. In maize, the transformation frequency (TF) using pat as a selectable marker is the lowest among the commonly used marker options including epsps, pmi or ppo. Low pat transformation efficiency can become a major bottleneck in our ability to efficiently produce large numbers of events, especially for large molecular stack vectors with multiple trait gene cassettes. The root cause of the lower efficiency of pat in maize is not well understood and it is possible that the causes are multifaceted, including maize genotype, pat marker cassette, trait gene combinations and selection system. In this work we have identified a new variant of pat gene through codon optimization that consistently produced a higher transformation frequency (> 2x) than an old version of the pat gene that has codons optimized for expression in dicot plants. The level of PAT protein in all 16 constructs was also found multifold higher (up to 40 fold) over that of the controls. All of the T0 low copy transgenic plants generated from the 16 different constructs showed excellent tolerance to ammonium glufosinate herbicide spray tests at 4x and 8x recommended field application rates (1x = 595 g active ingredient (ai)/hectare of ammonium glufosinate) in the greenhouse. |
| Databáze: | OpenAIRE |
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