【摘要】 为了验证稻瘟病新抗性基因Pita2的功能,利用长片段PCR技术从基因的供体品种PiNo.4中扩增了2个候选基因Pita2-1,Pita2-2,长度分别为6.1 kb和16.5 kb,电泳回收目的片段,然后利用双酶切分别克隆到植物表达载体pCAMBIA1300。对阳性克隆通过菌落PCR、酶切鉴定和测序分析,已成功地获得了2个候选基因的重组阳性克隆,为进一步研究Pita2基因的功能奠定了基础。更多还原

【Abstract】 Rice blast is one of the most devastating diseases of rice,and a serious threat to rice production.Exploring new rice blast resistance genes for breeding disease resistance strains is an important strategy.To study the function of two candidate genes Pita2-1 and Pita2-2,long-range PCR(LR-PCR) was employed to amplify the candidate genes from genomic DNA of the resistant cultivar PiNo.4.The appropriate products of Pita2-1(6.1 kb)and Pita2-2(16.5 kb) were purified and then ligated into the binary vector pCAMBIA1300 respectively.The positive clones were confirmed by colony PCR,restriction endonuclease digestion and sequencing,thus laid a foundation for dissecting the function of gene Pita2.更多还原