摘要
潮霉素磷酸转移酶基因(Hygromycin B phosphotransferase gene,HPT或HPH)是一个十分有效的用于植物的选择标记基因,水稻的转基因研究一般都以该基因为标记及检测基因。本研究以转基因水稻的T2代为材料,研究了苗期、分蘖期的完全伸展叶和乳熟期不同部位的离体叶片对潮霉素的抗性反应,同步提取相应叶片的DNA与RNA进行HPT基因的PCR、RTPCR与Northern blot鉴定,结果表明两者符合率达到97.46%,可见利用检测离体叶片对潮霉素敏感性的方法来筛选转基因水稻具有一定的快捷性与可靠性。
Hygromycin Phosphtobamfemse gene （HPT） is an effective selectable marker gene in rice transformation. We usually ues this gene as the marker and detection gene in the rice transgenic research. We chose two methods：one was the homomycin resistance leaves assay, which was choosing T2 generation of the transgenic rice as the material in this research, and have discussed the resistance reaction of the full expanded leaves at seedling and tillering stages and leaves from different parts of plants at milk stage to homomycin; the other one was molecular method, DNA and RNA was extracted with these leaves, PCR amplification, RT-PCR amplification and Northern blot with HPT gene. The results indicated that these two methods had 97.46% approached. So we can conclude that it is rapid, high feasibility and reliability to filter the transgenic rice plant by means of detecting the resistance reaction of the in virto leaves to the homomycin.
出处
《黑龙江八一农垦大学学报》
2010年第1期45-48,57,共5页
journal of heilongjiang bayi agricultural university
基金
黑龙江省教育厅课题(11511248)
作者简介
殷奎德（1964-），男，教授，沈阳农业大学博士研究生毕业，现主要从事生物化学与分子生物学的教学与科研工作。
