转ATP/ADP转运蛋白基因籼稻的初步研究

Preliminary studies on transgenic indica rice with four ATP/ADP transporter constructs

ATP/ADP转运蛋白(AATP)在质体膜上负责外源ATP的转入,直接关系到质体内ATP浓度,从而可能影响质体内相关的一些重要代谢途径(如淀粉合成).本研究利用4个转基因结构(水稻aatp正反义转基因结构和菊芋aatp正反义转基因结构),采用农杆菌介导法获得了113个转基因水稻克隆.PCR、Southern Blot验证和后代遗传分析表明,外源基因多数以较低拷贝数整合到水稻基因组中.RT-PCR相对定量检测分析表明,插入基因在水稻中高水平表达,且与内源aatp基因在转录水平上存在复杂的相互干扰,反义植株aatp的转录水平比正义株系的低.

ATP/ADP translocator protein （AATP） , which resides on the membrane of plastids, is charged of the counter-exchange of cytoplasm ATP with inner ADP, thus relating to several important ATP-depended metabolic pathways inside plastids （such as starch synthesis） , by its influence to the concentration of plastidic ATP. In this paper, four constructs of either sense or antisense cDNA （ a pair for os-aatp from Oryza sativa L. and the other for ht-aatp from Helianthus tuberosus L. ） were introduced into rice Minghui 86 （ O. sativa L. indica） , mediated by Agrobacterium. PCR, Southern Blot validation and genetic analysis on transgenic plants against Hygromycin suppression were performed to showed that exogenous genes were most integrated into rice genome with lower copies. At mRNA level, transcript of inserted genes and their interfering with that of endogenous aatp gene were confirmed by RT-PCR. AATP mRNA concentrations in all antisense lines were lower than that in their sense counterparts.