Cloning and Analysis of NBS-LRR Type Resistance Gene Analogues in Vigna radiata
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摘要: 根据已知的拟南芥SPR2基因、烟草抗花叶病毒N基因、亚麻L6基因等NBS-LRR抗病类基因(RGAs)保守序列设计引物,从野生绿豆基因组DNA中分离得到了1条515bp大小的目的片段,并命名为FGV-1(GenBank登录号为KF021265)。经BLAST分析表明,分离的绿豆RGAs与已报道的大豆、豇豆、芸豆等植物的RGAs有较高的同源性。通过对其编码的氨基酸序列分析表明,FGV-1基因翻译的氨基酸序列中含有植物抗病基因NBS-LRR区域的4个保守结构:GMGGVGKTT、LILDDVD、GSRVIVTTRD及GLPLA,推测FGV-1可能是绿豆NBS-LRR类抗性基因的核心区域。绿豆RGAs的分离将为进一步从绿豆中分离功能性抗病基因打下基础,也为研究绿豆种质资源的起源与进化提供借鉴。Abstract: Degenerate primers based on conserved sequences of the nucleotide binding site and 1eucine rich repeats(NBS-LRR)region from the cloned plant disease resistance genes were used to isolate resistance gene analogues(RGAs)from genomic DNA of Vigna radiata.The desired band(515bp)was cloned and sequenced.The band was named FGV-1and had been submitted to Genbank(accession number KF021265).Blastx analys showed highly homology with the reported resistance gene analogues Glycine max,Vigna unguiculata and Phaseolus vulgaris. The analysis of RGAs amino acid sequence structures suggested that FGV-1 was the core region of NBS-LRR resistance genes in Vigna radiata,which contained four conserved domains including GMGGVGKTT,LILDDVD, GSRVIVTTRD and GLPLAL.The RGAs isolated fromVigna radiata used in this study would provide the base for the further cloning of disease-resistance genes in Vigna radiata,and provide reference for the origin and evolution of Vigna radiata.
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Key words:
- Vigna radiate /
- NBS-LRR /
- RGA /
- Homology cloning /
- Sequence analysis
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