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作 者:丁锦平[1] 张庆琛[1] 魏理[1] 李成伟[1,2]
机构地区:[1]商丘师范学院生命科学系/植物与微生物互作重点实验室,河南商丘476000 [2]周口师范学院生命科学系/植物遗传与分子育种重点实验室,河南周口466001
出 处:《广东农业科学》2012年第7期50-51,57,共3页Guangdong Agricultural Sciences
基 金:转基因生物新品种培育重大专项子任务(2008ZX08010-004)
摘 要:根据已克隆的R基因的保守序列设计引物,在棉花不同抗性品种中克隆出500 bp左右大小片段,克隆测序并在NCBI上进行Blast分析,发现这些序列除一个序列外均与已经克隆的抗病基因类似物(RGAs)同源,且同源性高达98%以上。将这些从棉花中克隆的RGAs和植物中已克隆的几个典型的R基因序列建立进化树,棉花中的RGAs序列分别聚在3个不同的亚家族,可以通过后续的功能验证来确定这些序列片段的功能。The resistance gene analog mining of cotton using NBS conserved sequences.Primers were designed according to the sequences of conserved P-loop and transmembrane motifs.The primers were used to amplify the disease resistance gene analogs(RGAs) in cotton varieties with different resistance levels against pathogen.Several PCR products about 500 bp were obtained,and these PCR products were cloned and sequenced.Blast analysis showed that all of them,except for one,are homologous to other disease resistance gene analogues(RGAs) with the similarity more than 98%.N-J phylogenetic tree was constructed based on RGAs sequences of cotton and other resistance gene sequences sought from GenBank in NCBI.When the sequences of 15 TIR type RGAs were compared with known resistance gene(cf-5、L6、Mil-2、RPM1、PRS2、Bs2、Gpa2、I2C-1) sequences,the RGA sequences were clustered in three different subfamilies,the RGAs may further be used as molecular marker for screening of candidate disease-resistance genes in cotton.
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