玉米ZmSKIP基因的克隆转化

随着淡水资源的匮乏,干旱已成为影响玉米产量众多因素中最主要的非生物因素。而植物的耐旱性又属于数量遗传性状,这给广大遗传育种工作者带来了一个很大的难题。SKIP基因属于RNA调控基因,它参与调节生物体内多种RNA的剪切调控,进一步调整植物状态,应对逆境环境。利用BLAST搜索玉米基因组,得到相似度99%的玉米基因,命名为ZmSKIP。对该基因克隆和生物信息学分析,构建过量表达载体,使用原位转化法得到过表达该基因的转基因株。对转基因株进行耐旱性实验,并使用ELISA方法检测转基因株的ABA含量。结果显示,转基因株中ABA相对野生型上升明显,具有更高的耐旱性。初步推断ZmSKIP在玉米中可能调控相应的耐旱基因表达从而调节植物对逆境的适应能力。

Molecular cloning and transformation of ZmSKIP in maize

With the scarcity of fresh water resources, drought has become the most important abiotic factor affecting the yield of corn. The drought-tolerance in plants belongs to quantitative genetic traits and could bring a serious problem to the majority of genetic breeders. The SKIP belongs to the RNA regulation. It participates in the regulation of a variety of RNA splicing, adjusts the vegetative state, and copes with adverse environment. In this study, using BLAST search with the maize genome, we got a sequence of 99% similarity with the maize gene, which was named ZmSKIP. Cloning and bioinformatics analysis, constructing the overexpression vector of ZmSKIP, and using in planta transformation method, we obtained the overexpressing transgenic plants. We conducted drought tolerance experiments on the transgenic plants, and detected ABA content in the transgenic plants by ELISA. The results showed that the ABA content in the transgenic plant increased significantly compared to its wild-type one, which indicated the former with higher drought tolerance. We concluded that ZmSKIP in maize may regulate the expression of the corresponding gene, and regulate plant drought stress on adaptability correspondingly.