关键词: 桃, 磷脂酶Dα, 电子克隆, RT-PCR, Northern杂交

Abstract:

【Objective】 The aim of this study was to isolate and clone new gene coding for phospholipase Dα from Prunus persica. 【Method】 In silico cloning was used to isolate cDNA sequence of PLDα, RT-PCR and Northern blot analysis were used to test the sequence and to analyze its expression during storage at 4℃. 【Result】 The obtained sequence contained a complete ORF, from 172 bp to 2 604 bp, encoded 810 amino acids. Compared with other plants, such as tomato, strawberry and grape etc., it was found that the amino acids sequence encoded by phospholipase Dα gene was almost conserved. Semi-quantitative RT-PCR and Northern blot analysis indicated that PLDα could be induced by low temperature. 【Conclusion】 Full length PLDα gene was first isolated and characterized from peach fruit, suggesting it is an efficient technique for cloning novel genes by searching EST database with homologous gene of relative species. Transcription profile of PLDα mRNA has provided a scientific basis for further research on the relationship between PLDα and low temperature.

Key words: peach (Prunus persica), phospholipase Dα, in silico cloning, RT-PCR, Northern blot