Transgene integration complexity and expression stability following biolistic or Agrobacterium-mediated transformation of sugarcane.
作者： Hao Wu , Faisal Saeed Awan , Aloisio Vilarinho , Qianchun Zeng , Baskaran Kannan
DOI: 10.1007/S11627-015-9710-0
关键词: Southern blot 、 Biology 、 Transgene 、 Transformation efficiency 、 Molecular biology 、 genomic DNA 、 Gene expression 、 Expression cassette 、 Agrobacterium 、 Transformation (genetics)
摘要: Sugarcane (Saccharum spp. hybrids) accounts for 80% of the table sugar produced worldwide and is also a prime feedstock biofuel production. However, very few studies are available directly comparing Agrobacterium tumefaciens-mediated transfer T-DNA (AMT) biolistic minimal expression cassettes (BLT MC) regarding transgene complexity stability. In this study, transformation efficiency, integration pattern, level, stability were compared in commercially important sugarcane cultivar CP88-1762. A total 312 transgenic lines derived from AMT 250 BLT MC identified by PCR genomic DNA using nptII-specific primers. Lines analyzed with both qPCR (TaqMan®) NPTII ELISA to determine nptII copy number level. The results Southern blot analysis on selected highly correlated results. There no significant differences between two systems frequency single integration, or level when carried out same cassette, tissue culture, selection procedure 12 independent experiments. These findings suggested that provide suitable platforms generation elite events.
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