| 蝴蝶兰ACC合成酶基因片段的克隆及其反义表达载体的构建 |
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| 引用本文： | 崔波,蒋素华,马杰,张仙云,叶永忠. 蝴蝶兰ACC合成酶基因片段的克隆及其反义表达载体的构建[J]. 安徽农业科学, 2009, 37(34): 16781-16782 |
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| 作者姓名： | 崔波 蒋素华 马杰 张仙云 叶永忠 |
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| 作者单位： | 河南农业大学生命科学学院,河南郑州 450002;郑州师范高等专科学校生物工程研究所,河南郑州 450044;河南农业大学生命科学学院,河南郑州,450002;郑州师范高等专科学校生物工程研究所,河南郑州,450044 |
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| 摘 要： | ［目的］克隆蝴蝶兰ACC合成酶cDNA片段，构建其反义表达载体。［方法］根据已报道的蝴蝶兰ACC合成酶的基因序列，设计并合成了1对引物，通过RTPCR以蝴蝶兰总RNA中扩增出ACC合成酶的cDNA片段，将其连接到MD19T质粒载体上进行测序。用XbaⅠ和SacⅠ对重组质粒和pBI221酶切后连接，构建蝴蝶兰ACC合成酶的反义基因表达载体。［结果］获得了蝴蝶兰ACC合成酶的cDNA片段，测序结果显示，该片段与参考的已报道序列具有98.92%和76.36%的同源性。通过引入XbaⅠ和SacⅠ酶切位点，进行载体构建，构建了蝴蝶兰ACC合成酶的反义基因表达载体。［结论］成功构建了蝴蝶兰ACC合成酶的反义基因植物表达载体，为进一步研究其对蝴蝶兰花期和生长的影响打下了基础。
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| 关 键 词： | 蝴蝶兰 ACC合成酶 基因克隆 反义基因 植物表达载体构建 |
cDNA Cloning and Construction of Antisense Expression Vector of Phalaenopsis ACC Synthase |
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| CUI Bo et al. cDNA Cloning and Construction of Antisense Expression Vector of Phalaenopsis ACC Synthase[J]. Journal of Anhui Agricultural Sciences, 2009, 37(34): 16781-16782 |
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| Authors: | CUI Bo et al |
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| Affiliation: | CUI Bo et al(College of Life Science,Henan Agricultural University,Zhengzhou,Henan 450002) |
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| Abstract: | [Objective] The purpose of this reseach was to clone the cDNA fragment of ACC synthase(1-aminocyclopropane-1-carboxylate synthase,ACS)of phalaenopsis and to construct its antisense expression vector.[Method] A pair of specific primers were designed and synthesized according to the reported cDNA sequences of phalanopsis ACC synthase gene(GenBank:Z77854.1;AF004663.1).The partial cDNA of ACS was obtained from total RNA of phalaenopsis by RT-PCR.Then the cDNA fragment was linked to T-tailing pMD19-T vector for ... |
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| Keywords: | Phalaenopsis ACC synthase Gene cloning Antisense gene Construction of plant expression vector |
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