梨S基因芯片的试制及分子杂交条件的优化

园艺学报 ›› 2008, Vol. 35 ›› Issue (4): 475-486.

梨S基因芯片的试制及分子杂交条件的优化

江南^1,2, 谭晓风^1*,陈洪¹, 王秀利² ,张琳¹, 曾艳玲¹

（¹中南林业科技大学经济林育种与栽培国家林业局重点实验室,长沙410004;²湖南工业大学绿色包装与生物纳米技术应用湖南省重点实验室，湖南株洲 412008）

收稿日期:2007-10-19 修回日期:2008-03-02 出版日期:2008-04-25 发布日期:2008-04-25
通讯作者: 谭晓风

Study on Preparation of Prototype S-RNase Microarray and Molecule Hybridization Condition

JIANG Nan^1,2,TAN Xiao-feng¹,CHEN Hong¹,WANG Xiu-li²,ZHANG Lin¹, ZENG Yan-ling¹

(¹The Key Laboratory of Non-Wood Forest Product of State Forestry Administration, Central-South University of Forestry and Technology，Changsha 410004, China; ²Key Laboratory of Green Packaging and Biological Nanotechnology, Hunan University of Technology, Zhuzhou Hunan, 412008, China)

Received:2007-10-19 Revised:2008-03-02 Online:2008-04-25 Published:2008-04-25
Contact: TAN Xiao-feng

摘要/Abstract

摘要：

为研究寡核苷酸芯片在确定梨品种S基因型中的应用价值，根据梨自交不亲和基因的结构特点设计了部分S基因的检测探针，并制成寡核苷酸芯片；采用引物Cy3荧光标记法标记检测样品的PCR产物并进行杂交，以检测不同样品的S基因型。结果表明：通过对不同杂交温度、杂交时间等的探索获得了较好的反应条件并用制备的芯片检测了已知S基因型的梨样品，检测结果与各品种的已知基因型相符。证明寡核苷酸芯片检测梨品种的S基因型是一种切实可行的检测方法，若对芯片进一步完善，则今后在梨的自交不亲和性状的机理研究及梨自交不亲和性状的利用等方面将有着广阔的应用前景。

关键词: 梨, 自交不亲和性, S -RNase基因, 基因芯片, 分子杂交

Abstract:

To explore the application of oligonucleotide microarray in identification of pears S-genotype, some probes for S-RNase detection were designed according to the structure characteristic of self-incompatibility gene, and oligonucleotide microarray was prepared. The PCR products of S-RNase were amplified by Cy3-labeled primer and hybridized with the microarray in order to detect the S-genotype of pears. Optimized reaction conditions were gained by exploration of hybridization temperature and hybridization time. The microarray results of S-genotype of pear was similar with the RFLP and sequenced results. In conclusion:, the oligonucleotide microarray is an effective method in detecting S-genotype of pear. With the future improvement, it could be widely used in the study of self-incompatibility of pear.

Key words: pear, self-incompatibility, S-RNase gene, gene chip, hybridization

江南;谭晓风;陈洪;王秀利;张琳;曾艳玲. 梨S基因芯片的试制及分子杂交条件的优化[J]. 园艺学报, 2008, 35(4): 475-486.

JIANG Nan;;TAN Xiao-feng;CHEN Hong;WANG Xiu-li;ZHANG Lin;ZENG Yan-ling . Study on Preparation of Prototype S-RNase Microarray and Molecule Hybridization Condition [J]. ACTA HORTICULTURAE SINICA, 2008, 35(4): 475-486.

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