野生华东葡萄脂类相关质体蛋白基因VpPAP1的克隆与表达

Molecular Cloning and Expression of a Plastid Lipid-associated Protein VpPAP1 Related to Uncinula necator Infection in Vitispseudoreticulata

该研究以前期获得的葡萄cDNA全长文库为基础,采用RT-PCR技术克隆了中国野生华东葡萄‘白河-35-1’(Vitis pseudoreticulata‘Baihe-35-1’)相关质体蛋白基因,命名为VpPAP1(GenBank登录号JN624817)。序列分析表明,VpPAP1基因cDNA编码区全长为1 034bp,其中5′-UTR和3′-UTR区域分别为26bp和63bp,开放阅读框为945bp,编码314个氨基酸,分子量为34 169.37Da,等电点为7.76。葡萄叶片接种白粉菌[Uncinula necator(Schw.)Burr.]后,运用实时定量PCR技术分析VpPAP1基因的表达模式,结果表明VpPAP1基因受白粉菌诱导表达,在接种后24h达到峰值。该研究为进一步研究中国野生葡萄脂类相关质体蛋白基因的表达及其在葡萄白粉病互作中的功能分析提供了依据。

Based on our constructed cDNA full length library,a gene named VpPAPl（GenBank Accession NO. JN624817） encoding plastid lipid-associated protein was cloned and sequenced from leaves of Vitis pseudoreticulata ＇Baihe-35-1＇ by RT-PCR（Reverse Transcription PCR）. The sequence analysis indicated that the cDNA length was 1 034 bp,flanked by a 5＇-untranslated region of 26 bp and a 3＇-untranslated re- gion of 63 bp. 945 bp of the open reading frame encodes a polypeptide of 314 amino acid residues,protein molecular weight is 34 169.37 Da and theoretical isoelectric point 7.76. The expression profile of the Vp- PAP 1 was analyzed in V. pseudoreticulata infected Uncinula necator. Realtime-PCR analysis revealed that the VpPAP 1 gene was induced by U. necator in leaves, and reached the peak at 24 h, which indicated that it may involve in the resistance to U. necatori. The study will be helpful to study the function o{ plastid lipid- associated protein on interaction of U. necator and grapevine.