摘要
为了降低bFGF(basic fibroblast growth factor)的生产成本,结合植物生物反应器的优点,就bFGF在转基因苜蓿中的表达进行了探索。将bFGF插入植物表达载体pBII21中,获得了含有bFGF基因的植物表达pBIcbFGF,再将pBIcbFGF利用冻融法转到农杆菌中。利用农杆菌介导法将基因转化保定苜蓿,转基因苜蓿在TM-1培养基+20mg/L卡那霉素(Kan)+200mg/L特美汀(Tim)中诱导分化,在生根培养基中生根,获得再生植株。再生植株通过PCR检测、RT-PCR及Western blot证实外源基因已经在苜蓿中成功表达。获得含有目的蛋白的阳性植株。为苜蓿作为植物生物反应器生产bFGF奠定了理论及技术基础。
In order to reduce the production costs ofbFGF, we explored the expression ofbFGF in transgenic alfalfa. A plant expression vector pBIcbFGF was constructed by inserting bFGF gene into plasmid pBⅡ21, and trans-formed into Agrobacterium using Freeze-thaw method. Transformed into Baoding alfalfa using Agrobacterum-mediated transformation to obtain transgenie alfalfa. Genetically modified alfalfa at TM-1 medium ＋20 mg/L Kan＋ 200 mg/L Tim in the induction of differentiation, in the rooting medium induced root, to obtain regenerated plants. The successful expression of heterologous gene in alfalfa was confirmed by PCR, RT-PCR and Western blot. The purpose was to contain protein-positive plants. For alfalfa as a plant bioreactor for the production ofbFGF has laid a foundation for the theory and technology.
出处
《基因组学与应用生物学》
CAS
CSCD
北大核心
2009年第3期455-459,共5页
Genomics and Applied Biology
基金
国家863项目(20051113)资助
