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Chinese Horticulture Abstracts
2010,26
(4)
10-14
转基因甜橙定量PCR分析体系的建立
作 者：
严佳文;杨莉;袁飞荣;张家银;李娜;王晓君;邓子牛
单 位：
湖南省作物种质创新与资源利用重点实验室;湖南农业大学园艺园林学院
关键词：
pthA-nls基因;β-actin基因;甜橙;定量PCR
摘 要：
以转pthA-nls基因甜橙45d叶龄叶片、枳橙的根、茎、叶不同器官以及非转基因枳、枸橼、温州蜜柑、沙田柚的45d叶龄叶片为试材,采SYBR GreenⅠ荧光染料法,检验不同柑橘类型以及枳橙不同器官中β-actin基因表达的稳定性,并以β-actin基因作内参基因对pthA-nls基因pthA-nls基因甜橙中的表达量进行分析。结果表明:β-actin基因在柑橘的不同类型及同一类型的不同组织中表达均较恒定,可作为内参基因用于柑橘基因的定量PCR分析;同时,试验还应用建立的方法体系分析转pthA-nls基因甜橙,基本明确各转基因株系中目的基因表达水平。
译 名：
Expression of PthA-nls Gene in Sweet Orange Analyzed by Quantitative Polymerase Chain Reaction
作 者：
YAN Jia-wen YANG Li YUAN Fei-rong ZHANG Jia-yin LI Na WANG Xiao-jun DENG Zi-niu
关键词：
pthA-nls gene;;β-actin gene;;sweet orange;;quantitative PCR
摘 要：
45 days old leaves from transformed sweet orange with pthA-nls gene,different organs(roots,shoots,leaves and buds) from Citrange,and 45 days old leaves from untransformed Trifoliate,Satsuma Mandarin,Citron,Pomelo,Bingtang sweet orange,Citrange were tested.The stability of expression ofβ-actin gene was identified by SYBR GreenⅠfluorescent dye method.The results demonstrated the expression ofβ-actin gene in different varieties and tissues of citrus in a constant states.β-actin gene can be used as reference gene for quantitative PCR analysis in citrus and the relative expression level of pthA-nls gene in different transgenic sweet orange was detected withβ-actin as reference gene.
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