| 番茄S-腺苷蛋氨酸脱羧酶基因SlSAMDC1的克隆与序列分析 |
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| 引用本文： | 刘志勇,王孝宣,高建昌,国艳梅,杜永臣,叶雪凌.番茄S-腺苷蛋氨酸脱羧酶基因SlSAMDC1的克隆与序列分析[J].园艺学报,2008,35(8):1137-1146. |
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| 作者姓名： | 刘志勇 王孝宣 高建昌 国艳梅 杜永臣 叶雪凌 |
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| 作者单位： | （中国农业科学院蔬菜花卉研究所，北京100081;上海市农业科学院园艺所，上海201106;3首都师范大学生命科学学院，北京100037） |
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| 摘 要： | 以蔓陀罗S-腺苷蛋氨酸脱羧酶全长cDNA序列为信息探针, 筛选NCBI番茄EST数据库, 依据同源EST信息，经人工拼接、RT-PCR及RACE技术验证，获得了1个新的SAMDC基因家族成员，命名为SlSAMDC1(GenBank登录号：EF550528)，并利用染色体步移技术克隆了879 bp的上游调控区域。SlSAMDC1 cDNA序列全长1 847 bp, 5′-UTR和3′-UTR分别长523 bp、241 bp；存在3个ORF(微型uORF、小型uORF和主ORF)，主ORF编码360个氨基酸的SAMDC酶原；SlSAMDC1基因组序列全长3 648 bp，有3个内含子，均位于5′-UTR，所有内含子的剪切位点均符合真核生物“GT-AG”规则。SlSAMDC1基因与其它植物来源SAMDC基因同源性较高，与人、大肠杆菌以及酵母的同源性较低。表达谱分析发现，SlSAMDC1基因在番茄根、茎、叶、花蕾、果实等器官中均表达，果实中的表达量相对较高。生物信息学分析表明，SlSMDC1基因的上游调控序列存在多个顺式作用元件，如W-box、TATA-box、CAAT-box等。
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| 关 键 词： | 番茄 多胺 S-腺苷蛋氨酸脱羧酶 基因克隆 |
| 收稿时间： | 2008-3-5 |
| 修稿时间： | 2008-6-18 |
Cloning and Sequence Analysis of a S-Adenosylmethionine decarboxylase gene (SlSAMDC1) in tomato |
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| LIU Zhi-yong,WAND Xiao-xuan,GAO Jian-chang,GUO Yan-mei,DU Yong-chen,YE Xue-ling.Cloning and Sequence Analysis of a S-Adenosylmethionine decarboxylase gene (SlSAMDC1) in tomato[J].Acta Horticulturae Sinica,2008,35(8):1137-1146. |
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| Authors: | LIU Zhi-yong WAND Xiao-xuan GAO Jian-chang GUO Yan-mei DU Yong-chen YE Xue-ling |
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| Institution: | (Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081, China;Horticultural Research Institute, Shanghai Academy of Agricultural Sciences, Shanghai 201106, China;College of Life Science, Capital NormalUniversity, Beijing 100037, China) |
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| Abstract: | The SAMDC cDNA sequence from jimsonwead (GenBank Accession No: Y07768) was used as electronic probe to hybridize tomato ESTs in GenBank through blastn search. Based on the contig assembled from homologous ESTs, the full-lenghth cDNA of a new member of SAMDC family in tomato was cloned using RT-PCR and RACE，which is 1 847 bp in length with a 523 bp 5′-UTR and a 241 bp 3′-UTR. Corresponding DNA sequence is 3 648 bp, containing three introns in 5′-UTR, and all the cleave sites obey with the GT-AG rule. The gene was designated as SlSAMDC1 (GenBank Accession No: EF550528). SlSAMDC1 consists of three ORFs (tiny uORF, small uORF and main ORF), and the main ORF encodes 360 amino acids with a calculated molecular weight of 39kD. The comparison of deduced amino acid sequence revealed that SlSAMDC1 is highly homologous to other plant SAMDCs, but less similar to human and yeast′s. RT-PCR analysis showed that expression level of SlSAMDC1 was higher in fruit than in any other organs. The upstream regulatory sequence was cloned by genome walking. Further analysis showed that the upstream regulatory sequence contains several Cis-acting elements (eg. W-box, TATA-box, CAAT-box). |
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| Keywords: | Solanum lycopersicum tomato (Solanum lycopersicum polyamine S-Adenosylmethionine decarboxylase gene cloning |
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