摘要
微弹轰击法将外源DNA导入小麦(Triticum aestivum)未成熟胚和悬浮细胞系的完整细胞。用pCI GUS作报告基因,质粒DNA涂于钨粉微弹表层,用基因枪轰击使微弹穿透细胞壁进入小麦细胞。处理2天后,用X-glucuronide染色,对GUS基因产物的活性进行鉴定,GUS基因表达的细胞呈深蓝色的小点。小麦幼胚表面的蓝点最多可达40～50个,悬浮细胞系中GUS基因表达的细胞较少。试验表明微弹轰击法是小麦组织水平上进行外源基因导入的一个有效途径。试验对微弹轰击的有关参数及条件进行了讨论。
Foreign DNA had been introduced into intact cells of cultured immature embryos and suspension culture cells of wheat (Triticum aestivum),by microprojectile bombardment. Tungsten particles were coated with reporter gene pCI GUS, and passed through cell wall by using a particle gun.Two days after bombardment,activity of GUS product was detected by staining with X-glucuronide. The cells expressing β-glucuronidase gave instense blue coloured spots. Up to 40—50 blue spots showed on the surface of an immature embryo of wheat, but Jess spots in suspension cells. This approach presents an alternative method to transfer foreign genes into cells of wheat tissue.Some bombardment parameters and conditions were discussed.
出处
《生物工程学报》
CAS
CSCD
北大核心
1992年第4期339-341,共3页
Chinese Journal of Biotechnology
关键词
小麦
基因导入
微弹轰击
GUS基因
Wheat (Triticum aestivum)
gene transfer
transient gene expression
microprojectile bombardment
