石榴F3’H基因PCR反应体系的研究  

Establishment of PCR System for F3’H Gene of Pomegranate Flower

作　　者：陶吉寒[1] 招雪晴[1] 苑兆和[1] 尹燕雷[1] 冯立娟[1] 

机构地区：[1]山东省果树研究所,山东泰安271000

出　　处：《湖北农业科学》2013年第9期2168-2170,共3页Hubei Agricultural Sciences

基　　金：山东省自然基金项目(ZR2009DM010)

摘　　要：从石榴(Punica granatum L.)泰山红品种的花瓣中提取总RNA并反转录得到cDNA。设计简并引物对石榴类黄酮3’-羟化酶(F3’H)基因进行PCR扩增,筛选合适的引物及退火温度。结果表明,适合石榴F3’H基因扩增的正反向引物分别为5′-CGTNGAYGTBGTBGTBGCSKCVTC-3′,5′-TCHCCDGCWATG-GCCCAHAYRTT-3′。PCR反应程序为94℃预变性5 min;94℃变性40 s,54℃退火45 s,72℃延伸60 s,共35个循环;72℃保温10 min。Total RNA was extracted from petal of pomegranate（Pun/ca granatum L.） cultivar Taishanhong and used for reverse transcription to obtain cDNA. Degenerate primers were designed for PCR amplification of flavanone-3-hydroxylase（F3＇H） gene of pomegranate, suitable primers and annealing temperature were screened for the PCR system. The results showed that the forward and reverse primer for amplifying F3＇H gene was 5＇-CGTNGAYGTBGTBGTBGCSKCVTC-3＇, 5＇-TCHCCDGCWATG- GCCCAHAYRTY-3＇, respectively. The thermal profile consisted of 35 cycles of denaturation at 94 ℃ for 40 s, annealing at 54 ℃ for 45 s, and polymerization at 72 ℃ for 60 s, preceded by an initial denaturation step at 94 ℃ for 5 min and followed by a terminal extension at 72 ℃ for 10 min.

关 键 词：石榴(Punica granatum L ) F3’H基因 PCR 

分 类 号：S685.99[农业科学—观赏园艺]