本文選題：梨 + cDNA芯片��； 參考：《生物技術(shù)》2017年01期

【摘要】：[目的]優(yōu)化梨自交不親和基因(S-RNase或S基因)c DNA芯片雜交條件,利用芯片檢測梨品種S基因型。[方法]提取梨品種雌蕊RNA,Cy3標(biāo)記引物RT-PCR獲得S基因熒光標(biāo)記特異c DNA序列。設(shè)置不同雜交條件,用已知S基因型品種熒光標(biāo)記的PCR產(chǎn)物在不同條件下分別與芯片雜交,雜交信號分析芯片雜交效果。用芯片優(yōu)化雜交體系鑒定梨品種未知S基因型,DNA測序驗證芯片鑒定結(jié)果。[結(jié)果]芯片雜交最佳條件:雜交溫度42℃,雜交時間8~9 h,PCR純化產(chǎn)物終濃度為200 ng·μl-1。優(yōu)化雜交條件下芯片鑒定晚咸豐、秀水、麗江馬占梨1、湘菊、木通梨、甘甜、彌渡小紅梨、麗江大中古、金晶和彌渡火把等梨品種S基因型分別為:Pp S15Pp S52、Pp S4Pp S5、Pb S22Pp S37、Pp S1Pp S2、Pp S1Pp S3、Pp S13Pp S15、Pp S12Pb S42、Pb S21Pb S22、Pp S3Pp S60和Pp S5Pp S5。DNA測序驗證各品種所含S基因與芯片鑒定結(jié)果一致。[結(jié)論]梨自交不親和基因c DNA芯片優(yōu)化雜交條件后可準(zhǔn)確鑒定梨品種所含已鑒定的S基因資源。
[Abstract]:[objective] to optimize the conditions of S-RNase or S-gene DNA microarray hybridization of pear self-incompatibility gene, and to detect the S-genotypes of pear varieties by microarray. [methods] the C DNA sequence of S gene fluorescent marker was obtained from the primer RT-PCR labeled with Cy3 in pistil of pear cultivars. Under different hybridization conditions, the PCR products labeled with known S genotypes were hybridized with microarray under different conditions, and the hybridization signal was analyzed by chip hybridization. DNA sequencing of unknown S genotypes of pear cultivars identified by microarray optimized hybridization system was used to verify the results of microarray identification. [results] the optimal conditions for microarray hybridization were as follows: hybridization temperature 42 鈩，

本文編號：1797282