Factors influencing electroporation-mediated gene transfer to Stylosanthes guianensis (Aubl.) Sw. protoplasts

Genetics and Molecular Biology

V.m. Quecini¹  C.a. De Oliveira¹  A.c. Alves¹  M.l.c. Vieira¹ 

[1] ,USP ESALQ Departamento de GenéticaPiracicaba SP ,Brazil

关键词: direct gene transfer;    plant transformation;    forage legume;    Stylosanthes guianensis;   

DOI  :  10.1590/S1415-47572002000100014

来源: SciELO

【 摘 要 】

In order to develop a high-efficiency and reproducible transformation protocol for Stylosanthes guianensis we assessed the biological and physical parameters affecting plant electroporation protoplasts. Energy input, as combinations of electric field strengths discharged by different capacitors, electroporation buffer and DNA form were evaluated. Transformation efficiency was assayed in vivo as transient reporter gene expression, using the GFP-coding gene mgfp5 driven by a CaMV 35S constitutive promoter. Energy input and electric field strength had a critical influence on transgene expression with higher transformation levels being achieved with 250 V.cm-1 discharged by 900 and 1000 muF capacitors. Linear plasmid DNA, the absence of chloride and the presence of calcium ions also increased transient gene expression, albeit not significantly.

【 授权许可】

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