夏黑葡萄花及果实全长cDNA文库的构建及鉴定

Construction and Evaluation of cDNA Library of Flower and Fruit of Grapevine "cv. Summer Black"

构建葡萄花果全长cDNA文库是开展葡萄花及果实发育的分子机理研究的重要工作基础,有助于重要相关基因的克隆、功能分析、调控及其利用。以生产上广泛栽培、性状优良且极具代表性的夏黑葡萄为试材,应用优化的CreatorSMART cDNA Construction Kit技术,构建了夏黑葡萄花和果实的全长cDNA文库。文库的滴度为1.2×106pfu/mL,库容为6.0×106pfu/mL。从文库中随机挑取30个克隆进行菌落PCR鉴定的结果显示:插入片段大小为1.0～3.0kb,重组率为99%。研究结果表明,该葡萄全长cDNA文库具备较高的质量。

Grapevine is one economically important fruit crop in the world. Construction of the full-length cDNA library of grape flower and berry is the importantly fundamental work for the study on the molecular mechanism of the development of grape flower and berry,in which it can facilitate the cloning,characterization and utilization of the important genes involved. In this research,a full-length cDNA library of flower and berry of grapevine cv . ″Summer Black″,a well known grape cultivar in its high popularity in cultivation and great berry quality,was constructed with Creator SMART cDNA Construction Kit. This library has a capacity of 1. 2 × 10^6 clones,and its recombination efficiency was about 99% ,and the insert sizes were about 1. 0 - 3. 0 kb based on PCR analysis of 30 randomly chosen clones, suggesting that the cDNA library of grape flower and fruit has been successfully constructed for the use in further study.