College of Natural, Applied and Health Sciences, Wenzhou Kean University, Wenzhou, Zhejiang, China.
Department of Genetics and Biochemistry, Clemson University, Clemson, SC, USA.
Methods Mol Biol. 2020;2124:251-261. doi: 10.1007/978-1-0716-0356-7_13.
We describe a protocol for the establishment and preparation of creeping bentgrass (Agrostis stolonifera L.) cultivar "Penn A-4" embryonic calli, biolistic transformation, selection, and regeneration of transgenic plants. The embryonic callus is initiated from mature seeds, maintained by visual selection under the dissecting microscope and subjected to bombardment with plasmid DNA containing a bialaphos-resistance (bar) gene. PCR, Southern, and Northern blot analyses are used to confirm the transgene integration and expression.
我们描述了建立和准备匍匐翦股颖（Agrostis stolonifera L.）品种“Penn A-4”胚胎愈伤组织、生物弹道转化、转基因植物选择和再生的方案。胚胎愈伤组织由成熟种子起始，在解剖显微镜下通过目视选择进行维持，并进行含有草丁膦抗性（bar）基因的质粒 DNA 轰击。通过 PCR、Southern 和 Northern 印迹分析来确认转基因的整合和表达。
