基因枪轰击法将蜘蛛丝蛋白基因(ADF3)转入海岛棉的研究

Transformation of Spider Silk Protein Gene into the Apical Meristems of Gosspium Barbadens Via Particle Bambardment

用海岛棉茎尖作为基因枪转化的靶材料,建立了可重复的海岛棉转化系统。将海岛棉茎尖分生组织经过3～6周的诱导、继代培养生长后,可以形成足够量的再生植株。用含有Npt-Ⅱ基因和蜘蛛丝蛋白基因的质粒轰击新疆海岛棉4个品种茎尖分生组织,在含有70～100mg/l卡那霉素的培养基上筛选、预再生、再生及生根培养,获得的24株抗卡那霉素转基因再生植株。所获得的转基因植株经卡那霉素筛选,再生植株总DNA提取,Southern点杂交分子检测,初步证明有2株转化植株目的基因已整合到海岛棉基因组中,同时也已得到种子。研究为通过植物基因工程的方法以期改良棉花纤维品质提供了一条新的途径。

A reproducible and efficient transformation system for barbados cotton (Gossypium barbadens L.) was developed based on microprojectile bombardent. Sufficient plantlets of regeneration were produced within 3～6 weeks induced and culture from apical meristems. The apical merristems were bombarded with plasmid containing Npt-Ⅱ gene and spider silk prote in gene. Selection,pre-regeneraion,regeneration and rooting were carried out on media with 70～100mg/l kanamycin,and then 24 resist-Kanamycin transgenic plantlets were obtained. Transgenic plantlets were confirmed by Kanamycin-resistant selection and southern dot blotting analysis.The results preliminarily demonstrated the gene was integrated into the genome of transgenic cotton plants. The research offers a new method to improve fiber quality of cotton by plant gene engineering.