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[1]于妍,唐敬仙,姜威,等.大豆AK 基因的克隆与序列多样性分析[J].大豆科学,2016,35(04):574-580.[doi:10.11861/j.issn.1000-9841.2016.04.0574]
　YU Yan,TANG Jing-xian,JIANG Wei,et al.Molecular Cloning and Gene Diversity Analysis of Gene AK in Soybean[J].Soybean Science,2016,35(04):574-580.[doi:10.11861/j.issn.1000-9841.2016.04.0574]

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大豆AK 基因的克隆与序列多样性分析

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第35卷期数: 2016年04期页码: 574-580 栏目: 出版日期: 2016-07-20

Title:: Molecular Cloning and Gene Diversity Analysis of Gene AK in Soybean

作者:: 于妍¹; 唐敬仙¹; 姜威²; 蒋洪蔚³; 邱红梅⁴; 刘春燕³; 陈庆山²; 胡国华³; ( 1．长春科技学院生物食品学院，吉林长春130600; 2. 东北农业大学农学院，黑龙江哈尔滨150030; 3．黑龙江省农垦科研育种中心，黑龙江哈尔滨150090; 4. 吉林省农业科学院大豆研究所，吉林长春130033)

Author(s):: YU Yan1; TANG Jing-xian1; JIANG Wei2; JIANG Hong-wei3; QIU Hong-mei4; LIU Chun-yan3; CHEN Qingshan2; HU Guo-hua3; ( 1. College of Biology and Food，Changchun University of Science and Technology，Changchun 130600，China; 2. College of Agriculture，Northeast AgriculturalUniversity，Harbin 150030，China; 3. The Crop Ｒesearch and Breeding Center of Heilongjiang Land Ｒealamation，Harbin 150090，China;4. Soybean Ｒesearch Institute，Jilin Academy of Agricultural Sciences，Changchun 130033，China)

关键词:: 大豆; 天冬氨酸代谢; 天冬氨酸激酶( AK) ; 基因多样性

Keywords:: Soybean; Aspartic acid metabolism; Aspartate kinase( AK) ; Gene diversity

分类号:: S565. 1

DOI:: 10.11861/j.issn.1000-9841.2016.04.0574

文献标志码:: A

摘要:: 基于NCBI 网站下载获得的大豆AK 基因cDNA 序列设计特异引物，以大豆品种东农42 总ＲNA 为模板，通过ＲT-PCＲ获得约1 690 bp cDNA 片段，经序列比对认定为AK 基因序列。以19 个赖氨酸和蛋氨酸含量特殊的大豆品种为材料，采用相同方法在ＲNA 中进行扩增，并对产物测序，利用软件对所得序列进行分析。结果显示: 在不同的大豆品种中AK 基因的核酸序列存在两种变异方式，第一种是在336 bp 产生1 个T 碱基突变的基因序列，第二种是在1 369 ～ 1 374 bp 处出现“CAAGTG”6 个碱基插入的序列; 在蛋白质水平上，主要是在456 ～ 457 个氨基酸处存在A 和S两个氨基酸插入突变。AK 基因结构完整，其编码的蛋白产物具有AA_kinase、Carbamate kinase-like、Aspartate kinase 和ACT 保守结构域。本研究首次对大豆中AK 基因多样性进行了分析，为大豆天冬氨酸代谢途径其它相关酶基因的研究提供了理论依据，也为大豆AK 基因在植物基因工程等领域的研究和应用奠定了良好的基础。

Abstract:: AK gene was cloned from soybean，a cDNA product about 1 690 bp was amplified from the total ＲNA of soybeanleaves by reverse transcription PCＲ( ＲT-PCＲ) with specific primers designed with the full length sequence of AK gene fromNCBI． The cDNA of 19 varieties were amplified with the same way. The PCＲ products were cloned，sequenced，and analyzedwith bioinformatics software. The results showed that there were two mutant alleles of nucleotide sequences in different soybeanvarieties，the first was the sequence which had a nucleotide substitution，the second was the sequence which had an insertionof 6 nucleotides between 1 369 and 1 374 bp. There were only one mutant allele of amino acid sequences in protein level，which was an insertion of 2 amino acids at 456 － 457 amino acid residue. The structure of AK was completed，the conserveddomains of AA_kinase，Carbamate kinase-like，Aspartate kinase，and ACT were found in protein of AK. The gene diversity analysisof gene AK in soybean was studied for the first time in this research. The results provided a theoretical basis for thestudy of other related enzymes in the aspartate pathway of soybean，and also laid a good foundation for the research and applicationof AK gene in plant genetic engineering and other fields.

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备注/Memo

基金项目: 长春科技学院校内科研启动基金项目; 黑龙江省自然科学基金重点项目( ZD201213) ; 教育部新世纪优秀人才支持计划( NECT-1207-01) 。第一作者简介: 于妍( 1984-) ，女，硕士，讲师，主要从事大豆遗传育种研究。E-mail: 35789362@ qq． com。通讯作者: 陈庆山( 1973-) ，教授，博导，主要从事大豆遗传育种研究。E-mail: qshchen@126． com;胡国华( 1951-) ，研究员，博导，主要从事大豆遗传育种研究。E-mail: hugh757@ vip． 163． com。

更新日期/Last Update: 2016-08-22