稻瘟病新抗性基因Pita2候选基因的表达载体构建  

Construction of Expression Vector for Candidate Genes of New Rice Blast Resistance Gene Pita2

作　　者：张骥诚[1] 张向明[1] 王春台[1] 刘学群[1] 徐鑫[1] 刘新琼[1] 

机构地区：[1]中南民族大学生命科学学院生物技术国家民委重点实验室,武汉430074

出　　处：《湖北农业科学》2011年第10期2133-2135,共3页Hubei Agricultural Sciences

基　　金：国家自然科学基金项目(30971563);湖北省自然科学基金项目(2009CDA125;2010CDB02006)

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。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.

关 键 词：稻瘟病 候选抗性基因 长片段PCR技术 表达载体 

分 类 号：Q786[生物学—分子生物学]