期刊论文

中文

生物技术

1004-311X

2017

27

1

58-64+70

10.16519/j.cnki.1004-311x.2017.01.0010

国家自然科学基金项目（“基于基因芯片技术的砂梨品种S基因型鉴定及S基因资源挖掘与进化分析” No.31272124）

本校为第一机构

[目的]优化梨自交不亲和基因（S-RNase或S基因）c DNA芯片杂交条件,利用芯片检测梨品种S基因型。[方法]提取梨品种雌蕊RNA,Cy3标记引物RT-PCR获得S基因荧光标记特异c DNA序列。设置不同杂交条件,用已知S基因型品种荧光标记的PCR产物在不同条件下分别与芯片杂交,杂交信号分析芯片杂交效果。用芯片优化杂交体系鉴定梨品种未知S基因型,DNA测序验证芯片鉴定结果。[结果]芯片杂交最佳条件：杂交温度42℃,杂交时间8~9 h,PCR纯化产物终浓度为200 ng·μl-1。优化杂交条件下芯片鉴定晚咸丰、秀水、丽江马占梨1、湘菊、木通梨、甘甜、弥渡小红梨、丽江大中古、金晶和弥渡火把等梨品种S基因型分别为：Pp S...

[Objective]To optimize the hybridization condition of self-incompatibility gene（S-RNase or S gene） c DNA microarray and by the chip to identify S-genetypes of pear cultivars. [Methods]Total RNA was extracted from pear pistils,with which fluorescently-labeled specific c DNA sequences of S-RNase were amplified by RT-PCR amplification using Cy3-labeled specific primers. The hybridization effects of that the purified PCR products of pear cultivars with known S-genotype were hybridized with the c DNA microarray in different condition were analysed...