烟草叶片表面抗性蛋白T-phylloplanin基因的克隆
Clone of Resistant Protein-related T-phylloplanin Gene from Tobacco Leaf Surface
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摘要: 从烟草突变体T-cldf叶片表达基因的SSH文库中,筛选到一个与真菌抗性相关的表达序列标签P1T10(GenBank Access No.EB102896)。以P1T10序列为模板设计引物,用末端快速扩增(RACE)技术扩增得到T-Phylloplanin的cDNA全长(GenBank Access No.DQ451214)。利用DNASTAR软件的EditSeq程序(Version 5.01)分析T-Phylloplanin基因,显示它包含一个完整的开放读码框,编码一个含有110个氨基酸、等电点为7.74、分子量为11.3 kD的小分子量蛋白(GenBank Access No.ABE03627)。通过BLAST比对发现它与烟草叶片的一个表面抗性蛋白全长基因(GenBank Access No.AY705384)的同源性为93%。通过半定量RT-PCR分析表明,T-Phylloplanin在烟草突变体T-cldf叶片中的表达量高于烟草K346。Abstract: sen University,Guangzhou 510275,China P1T10(GenBank Access No.EB102896),an expression sequence tag(EST) which related to fungal-resistant was selected from a SSH(Suppression Subtractive Hybridization) pool.The complete cDNA of T-Phylloplanin gene(GenBank Access No.DQ451214) was gained by RACE(Rapid Amplification of cDNA End) with primers designed according to the sequence of P1T10.With the EditSeq program(version 5.01) of DNA STAR software T-Phylloplanin was analyzed,the results indicated that it contained a complete ORF(open reading frame) and a protein containing 110 amino acids was encoded whose weight was 11.3kD and pI(isoelectric point) was 7.74(GenBank Access No.ABE03627).The homology analysis by BLAST indicated that the similarity was 93% between T-phylloplanin and the complete cDNA gene(GenBank Access No.AY705384) of a resistant protein from tobacco leaf surface.Semi-quantitative RT-PCR analysis indicated that the T-phylloplanin expression was higher in mutant T-cldf leaf than in tobacco K346.
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