文献类型：期刊文献

中文题名：ACC合酶cDNA克隆及其对美洲黑杨体内乙烯产生的反义抑制

英文题名：CLONING OF ACC SYNTHASE cDNA AND ITS INHIBITION OF ETHYLENE SYNTHESIS BY ITS ANTISENSE RNA IN TRANSGENIC POPULUS DELTOIDES

作者：李明亮[1] 韩一凡[1] 邱德有[1] 李凝[2] 田颖川[3]

第一作者：李明亮

机构：[1]中国林业科学研究院林业研究所;[2]香港科技大学生物学系;[3]中国科学院微生物研究所

年份：1999

卷号：35

期号：3

起止页码：10-15

中文期刊名：林业科学

外文期刊名：SCIENTIA SILVAE SINICAE

收录：CSTPCD;;Scopus;北大核心:【北大核心1996】；CSCD:【CSCD2011_2012】；

基金：国家自然科学基金

语种：中文

中文关键词：美洲黑杨;黑杨;ACC合酶;cDNA;乙烯;反义抑制

外文关键词：ACC (l aminocyclopropane l carboxylate)synthase, Gene, Antisense RNA, Populus deltoides

分类号：S792.119

摘要：利用ＲＴＰＣＲ技术克隆了大豆ＡＣＣ合酶基因ＧＭＣＡＣＣＳＩ编码区１５ｋｂ的ｃＤＮＡ片段，经酶切图谱分析和序列分析鉴定后，反向插入到２元载体ｐＢｉｎ４３８中，构建了表达ＡＣＣ合酶反义ＲＮＡ的植物表达载体，转化农杆菌后用该农杆菌侵染美洲黑杨叶片，在含卡那霉素的ＭＳ培养基上选择转化子和植株再生，通过ＰＣＲ检测从抗卡那植株中选到１５株转基因植株，Ｓｏｕｔｈｅｒｎ杂交分析初步确证了外源基因是以单拷贝插入到杨树基因组ＤＮＡ中；对杨树幼苗乙烯释放的测定结果表明转基因杨树幼苗的乙烯释放量为对照的２２％。
A 1.5kb fragment of ACC synthase cDNA fragment prepared from total RNA of soybean was amplified by polymerase chain reaction(PCR) and cloned into pGEM T vector.The cloned ACC synthase gene was further inserted into a binary vector,pBin438,in an inverted orientation between the CaMV 35S promoter and Nos 3′termination sequence (pBACS).Transgenic poplar plants were obtained by regeneration of Agrobacterium mediated transformation of leaves of Populus deltoides. PCR and Southern blotting analyses confirmed the integration of a single antisense ACC synthase gene in transformed poplar genome.The results form reverse transcription PCR (RT PCR) of RNAs isolated from transgenic poplar leaves confimed that the antisense RNA of ACC synthase presented in these transgenic plants.The amount of ethylene released from transgenic poplar was reduced significantly to about 22% of that released from non transformed control poplars.