根据抗病基因核苷酸结合位点(Nuc leotide b ind ing site,NBS)设计简并性引物,从陆地棉cDNA中进行RT-PCR扩增。获得含NBS保守域的EST,进一步用RACE技术和TAIL PCR技术获得其中1个EST的全长基因序列,并获得GHNBS基因的5′调控序列。此...根据抗病基因核苷酸结合位点(Nuc leotide b ind ing site,NBS)设计简并性引物,从陆地棉cDNA中进行RT-PCR扩增。获得含NBS保守域的EST,进一步用RACE技术和TAIL PCR技术获得其中1个EST的全长基因序列,并获得GHNBS基因的5′调控序列。此基因被命名为GHNBS。该基因的编码区长2 583 bp,编码861个氨基酸,GHNBS编码的氨基酸序列与拟南芥R基因具有28%的同源性。GHNBS与拟南芥的其他几个NBS-LRR基因比较发现,它们在保守区外的相似性相当低。Southern杂交和网上数据库搜索分析都表明GHNBS是1个寡拷贝基因。通过半定量RT-PCR分析发现GHNBS在棉花的蕾、花瓣、韧皮部、根及叶中均有表达且在根、叶中表达量比其它部位强,在木质部基本不表达。展开更多
A full set of disease resistance(R) candidate genes encoding nucleotide-binding sites(NBS) in a complete genome of Populus trichocarpa was identified and characterized by structural diversities,physical positions,phyl...A full set of disease resistance(R) candidate genes encoding nucleotide-binding sites(NBS) in a complete genome of Populus trichocarpa was identified and characterized by structural diversities,physical positions,phylogenetic relationships.Based on structures of N-terminal motif and leucine-rich repeat domains motif,we found 381 NBS-coding sequences with 122 non-regular NBS genes and 259 regular NBS genes that were further classified into 13 types such as TNL,CNL,NL,XNL,TN and other minor types.Meanwhile 81.9% of the NBS genes were distributed in cluster,and 81.8% of the cluster genes had duplicates.The results showed that there were many duplicate phenomenon occurred in the evolution of disease resistance genes of P.trichocarpa.After analysis of NBS standard phylogenetic tree in the genome of P.trichocarpa,the structure of tree exhibited a star topology,and the regular NBS genes were classified into 68 groups by less than 30% amino acid sequence diversity in each domain.展开更多
文摘根据抗病基因核苷酸结合位点(Nuc leotide b ind ing site,NBS)设计简并性引物,从陆地棉cDNA中进行RT-PCR扩增。获得含NBS保守域的EST,进一步用RACE技术和TAIL PCR技术获得其中1个EST的全长基因序列,并获得GHNBS基因的5′调控序列。此基因被命名为GHNBS。该基因的编码区长2 583 bp,编码861个氨基酸,GHNBS编码的氨基酸序列与拟南芥R基因具有28%的同源性。GHNBS与拟南芥的其他几个NBS-LRR基因比较发现,它们在保守区外的相似性相当低。Southern杂交和网上数据库搜索分析都表明GHNBS是1个寡拷贝基因。通过半定量RT-PCR分析发现GHNBS在棉花的蕾、花瓣、韧皮部、根及叶中均有表达且在根、叶中表达量比其它部位强,在木质部基本不表达。
文摘A full set of disease resistance(R) candidate genes encoding nucleotide-binding sites(NBS) in a complete genome of Populus trichocarpa was identified and characterized by structural diversities,physical positions,phylogenetic relationships.Based on structures of N-terminal motif and leucine-rich repeat domains motif,we found 381 NBS-coding sequences with 122 non-regular NBS genes and 259 regular NBS genes that were further classified into 13 types such as TNL,CNL,NL,XNL,TN and other minor types.Meanwhile 81.9% of the NBS genes were distributed in cluster,and 81.8% of the cluster genes had duplicates.The results showed that there were many duplicate phenomenon occurred in the evolution of disease resistance genes of P.trichocarpa.After analysis of NBS standard phylogenetic tree in the genome of P.trichocarpa,the structure of tree exhibited a star topology,and the regular NBS genes were classified into 68 groups by less than 30% amino acid sequence diversity in each domain.
