粒子轰击辅助肽介导的基因转移用于高效瞬时测定。
Particle bombardment-assisted peptide-mediated gene transfer for highly efficient transient assay.
机构信息
Faculty of Agriculture, Takasaki University of Health and Welfare, Takasaki-shi, Gunma, Japan.
Faculty of Agriculture, Kyushu University, Fukuoka, Japan.
出版信息
BMC Res Notes. 2023 Apr 6;16(1):46. doi: 10.1186/s13104-023-06320-3.
OBJECTIVE
A centrifugation-assisted peptide-mediated gene transfer (CAPT) method was recently developed as an efficient system for gene delivery into plant cells. However, the gene transfer efficiency of CAPT into plant cells was not entirely satisfactory for detecting transient expression of a transgene driven into mitochondria. Here, we report a new gene delivery system using a method called particle bombardment-assisted peptide-mediated gene transfer (PBPT).
RESULTS
We investigated various parameters of the PBPT method to increase transient gene expression efficiency in Brassica campestris. The optimal conditions for PBPT were a single bombardment with gold particles coated with a DNA‒peptide complex (6 µg of DNA and 2 µg of peptide) at an acceleration pressure of 5 kg/cm and a target distance of 12.5 cm. Moreover, bombardment under the optimal conditions successfully transferred the transgene into the cells of other plant species, namely B. juncea and tomato. Thus, we developed a PBPT method for highly efficient delivery of a DNA‒peptide complex into plant mitochondria.
目的
最近开发了一种离心辅助肽介导的基因转移 (CAPT) 方法，作为将基因递送至植物细胞的有效系统。然而，对于检测驱动到线粒体的转基因的瞬时表达，CAPT 进入植物细胞的基因转移效率并不完全令人满意。在这里，我们报告了一种使用称为粒子轰击辅助肽介导的基因转移 (PBPT) 的新基因传递系统。
结果
我们研究了 PBPT 方法的各种参数，以提高 Brassica campestris 中转基因的瞬时表达效率。PBPT 的最佳条件是在加速压力为 5 kg/cm 和目标距离为 12.5 cm 的情况下，用涂有 DNA-肽复合物（6 µg DNA 和 2 µg 肽）的金颗粒进行单次轰击。此外，在最佳条件下的轰击成功地将转基因转入其他植物物种，即 B. juncea 和番茄的细胞中。因此，我们开发了一种 PBPT 方法，用于高效地将 DNA-肽复合物递送至植物线粒体中。
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