Schöpke C, Taylor N, Cárcamo R, Konan N K, Marmey P, Henshaw G G, Beachy R N, Fauquet C
International Laboratory for Tropical Agricultural Biotechnology (ILTAB/ORSTOM-TSRI), Scripps Research Institute, La Jolla, CA 92037, USA.
Nat Biotechnol. 1996 Jun;14(6):731-5. doi: 10.1038/nbt0696-731.
A protocol was established for the introduction of DNA into embryogenic suspension-derived tissues of cassava via microparticle bombardment, for the selection of genetically transformed cells, and for the regeneration of fully transgenic plants from these cells. The plasmid DNA used for bombardment contained a gene encoding neomycin phosphotransferase (nptII) and a gene encoding beta-glucuronidase (uidA). Selection of bombarded tissue with paromomycin resulted in the establishment of putative transgenic embryogenic calli. In most of these calli, beta-glucuronidase was detected histochemically. Molecular analysis of paromomycin-resistant embryogenic calli and of plants regenerated from these calli, confirmed the stable integration of bombarded DNA into the cassava genome.
建立了一种通过微粒轰击将DNA导入木薯胚性悬浮培养衍生组织的方案,用于选择基因转化细胞,并从这些细胞再生出完全转基因的植株。用于轰击的质粒DNA包含一个编码新霉素磷酸转移酶(nptII)的基因和一个编码β-葡萄糖醛酸酶(uidA)的基因。用巴龙霉素筛选轰击后的组织,建立了推定的转基因胚性愈伤组织。在大多数这些愈伤组织中,通过组织化学方法检测到了β-葡萄糖醛酸酶。对巴龙霉素抗性胚性愈伤组织以及从这些愈伤组织再生的植株进行分子分析,证实了轰击的DNA稳定整合到木薯基因组中。