水稻STN8激酶基因反义表达载体的构建及遗传转化

Construction and transformation of STN8 gene antisense-expression vector in rice

采用RT-PCR(reverse transcription polymerase chain reaction)技术从水稻中克隆获得光系统Ⅱ(photosnythesis systemⅡ,PSⅡ)蛋白激酶STN8基因片段.利用反义抑制技术,将STN8部分序列片段反向插入到植物表达载体pHB中,构建了植物反义表达载体pHBantis tn8.将重组载体转化农杆菌EHA105感受态细胞中,利用农杆菌介导法将其转入水稻.提取转基因植株基因组DNA,通过PCR检测转基因水稻中潮霉素基因的表达,初步筛选获得了阳性植株.转基因阳性苗的获得为进一步研究水稻PSⅡ蛋白激酶STN8的功能奠定了基础.

PSⅡ （photosnythesis system Ⅱ, PS Ⅱ） protein kinase STN8 gene fragment was obtained from Nipponbare using RT-PCR （reverse transcription polymerase chain reaction） method in this study. By antisense suppression technique stn8 was reversely inserted into plasmid pHB and the plant anti-expression vector pHBanti stn8 was constructed. After the recombinant vector was transformed into Agrobacteriurn EHA105, it was transformed into Nipponbare by Agrobacterium-mediated transformation. The total DNA of the transgenic plants was extracted and the positive transgenic plants were preliminarily confirmed by detecting the expression of Hygromycin using PCR. Altogether, the obtainment of positive transgenie plants facilitates the function analysis of STN8 in rice.