本研究利用质脂体介导法将构建好的家蚕转基因载体piggyBacA3EG转染家蚕培养胚胎细胞SWAU1-BraE,转染后48h在荧光显微镜下观察到大量的发绿色荧光的细胞,200h后,仍有部分细胞发绿色荧光。通过细胞爬片观察发现,绿色荧光是由细胞的胞质部分发出的,细胞核不发光。经RT—PCR检测,能在转染细胞cDNA中扩增出EGFP片段,表明该表达载体构建正确,且能在细胞水平进行瞬时表达,可以用于家蚕转基因的下一步研究。
By using the constructed vector pBacA3EG,. We transfected it - BmE and observed the green fluorogenic ceils after 48 hours later. into silkworm ceil line SWAU1 Through testing at the molecular level, the EGFP fragment was amplified from the cDNA of the transfected cell. The results indicated that the vector was constructed correctly and could express in the silkworm cell line, and can be applied in the transgenic studies of the silkworm.