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作 者:张永芳 李富恒[1] 吕怡杰 邱丽娟[3] ZHANG Yong-fang;LI Fu-heng;LYU Yi-jie;QIU Li-juan(School of Life Science,Northeast Agricultural University,Harbin 150030,China;School of Life Science,Shanxi Datong University,Datong 037009,China;Institute of Crop Sciences,Chinese Academy of Agricultural Science/The National Key Facility for Crop Gene Resources and Genetic Improvement/Key Laboratory of Germplasm&Biotechnology(Ministry of Agriculture and Rural Affairs),Beijing 100081,China)
机构地区:[1]东北农业大学生命科学学院,黑龙江哈尔滨150030 [2]山西大同大学生命科学学院,山西大同037009 [3]中国农业科学院作物科学研究所/国家农作物基因资源与遗传改良重大科学工程/农业农村部种质资源与生物技术重点开放实验室,北京100081
出 处:《大豆科学》2021年第1期11-20,共10页Soybean Science
基 金:“十三五”重点研发计划课题(2016YFD0100201)。
摘 要:为研究大豆中甜菜碱醛脱氢酶(BADH)在大豆中的分子机理及生物多样性,通过基因序列同源比对搜索到两个与水稻同源的大豆BADH基因,利用Phytozome、Netphosk 3.0 Server等网站及数据库对其进行生物信息学分析,结合已有的重测序数据分析1 598份大豆种质中BADH基因序列的多态性,并测定突变体的2AP含量。结果表明:大豆中有两个与水稻BADH基因同源性较高的基因序列Gm06g186300和Gm 05g0330550,基因全长分别为3 959和6 008 bp,编码蛋白均属于稳定蛋白质,二者同源性高达90%。两个蛋白的氨基酸数目、分子量和等电点等非常相似;亲水性和磷酸化位点分布较一致,均无跨膜结构域,在细胞核出现的可能性最大,且二者进化关系极其相近;α螺旋和无规则卷曲是两者的主要成分,BADH1和BADH2有1个共同的结构域Pfam-Aldedh,且分布大致相同。对1 598份大豆种质BADH基因序列突变位点进行分析,有29份材料BADH2基因发生6个碱基序列突变导致了移码突变,这种突变可能与大豆的芳香性表型有关。对BADH1及BADH2在大豆不同部位的基因表达量分析发现各部位的表达量相差很大,均以根部表达量最高,可能与根部较高的抗旱能力有关。In order to study the molecular mechanism and biodiversity of BADH in soybean,two soybean BADH genes with the same origin as rice were identified by sequence homology analysis blast function of NCBI website. Bioinformatics software Phytozome, Netphosk 3.0 Server and other websites and databases were further utilized to analyze the protein characteristics and structural characteristics of these two genes.The polymorphism of BADH gene sequence in 1 598 soybean germplasm was analyzed with resequencing data and the content of 2 AP was determined. The results showed that there were two gene sequences Gm06 g186300 and Gm05 G0330550 with high homology with rice BADH gene in soybean, the total length of which were 3 959 and 6 008 bp, respectively.It was found that the encoded proteins were stable, and the homology was as high as 90%. BADH1 and BADH2 were very similar in the number of amino acids, molecular weight, isoelectric point and so on. The distribution of hydrophilic and phosphorylation sites of BADH1 and BADH2 were consistent, and there was no transmembrane domain in the nucleus and the evolutionary relationship between them was very similar. According to the prediction scores of the secondary structures(α Helix, β angle, irregular curl, extended chain)of BADH1 and BADH2 amino acid sequences, it can be seen that α helix and random coil were the main components of them. BADH1 and BADH2 had only one common domain Pfam aldedh, and the distribution was roughly the same.The mutation sites of BADH Gene of 1 598 soybean germplasms were analyzed. Frame shift mutation was found in 6 base sites of BADH2 gene in 29 soybean germplasms. The aromatic component 2 AP of soybean germplasm with gene mutation was identified and the nucleotide sequence of the locus may be related to the aromatic phenotype of soybean.BADH1 and BADH2 gene expression analysis in different parts of soybean showed that the expression levels of each part were very different, and the highest expression level was found in the root, which may be related t
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