[1]聂腾坤,赵琳,李海燕,等.大豆-MPBQ-MT-基因的克隆与生物信息学分析[J].大豆科学,2016,35(01):39-45.[doi:10.11861/j.issn.1000-9841.2016.01.0039]
　NIE Teng-kun,ZHAO Lin,LI Hai-yan,et al.Cloning and Bioinformatics Analysis of MPBQ-MT Gene in Soybean[J].Soybean Science,2016,35(01):39-45.[doi:10.11861/j.issn.1000-9841.2016.01.0039]
点击复制

大豆-MPBQ-MT-基因的克隆与生物信息学分析

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第35卷 期数: 2016年01期 页码: 39-45 栏目: 出版日期: 2016-01-20
Title:
Cloning and Bioinformatics Analysis of MPBQ-MT Gene in Soybean
作者:
聂腾坤; 赵琳; 李海燕; 李文滨
东北农业大学 大豆生物学教育部重点实验室/农业部东北大豆生物学与遗传育种重点实验室，黑龙江 哈尔滨 150030
Author(s):
NIE Teng-kun; ZHAO Lin; LI Hai-yan; LI Wen-bin
Key Laboratory of Soybean Biology in Chinese Ministry of Education/Key Laboratory of Soybean Biology and Breeding Genetics of Chinese Agriculture Ministry，Northeast Agricultural University，Harbin 150030，China
关键词:
大豆; 维生素E; MPBQ-MT-基因; 生物信息学分析
Keywords:
Soybean; Vitamin E; MPBQ-MT gene; Bioinformatics analysis
DOI:
10.11861/j.issn.1000-9841.2016.01.0039
文献标志码:
A
摘要:
以大豆品种合丰25和Bayfield为材料，克隆大豆2-甲基-6-植基-1,4-苯醌甲基转移酶（MPBQMTase）基因，获得1 029 bp的MPBQ-MT-基因cDNA序列。通过序列比对发现10个大豆MPBQ-MT-基因cDNA区内的潜在SNP位点。采用生物信息学方法分析MPBQ-MT-基因编码的氨基酸序列和蛋白质结构。结果表明：MPBQ-MT-基因CDS区共编码342个氨基酸，其中缬氨酸（Val）含量最多，占该基因编码氨基酸总数的8.8%；半胱氨酸（Cys）含量最少，占该基因编码氨基酸总数的1.2%；MPBQ-MT蛋白N端包含由58个氨基酸组成的叶绿体转运肽，为亲水性蛋白质。合丰25与Bayfield的MPBQ-MT蛋白质二级结构有所不同，2个品种的MPBQ-MT蛋白质中α-螺旋分别约占25.44%和27.49%，β-转角分别约占9.36%和9.06%，无规则卷曲分别约占40.35%和38.60%。2个品种的MPBQ-MT蛋白质中片层结构均约占24.85%。两个品种的MPBQ-MT蛋白质均有1个S-腺苷基甲硫氨酸结合位点。对MPBQ-MT蛋白三级结构进行预测，发现去掉转运肽后，MPBQ-MT蛋白质三级结构主要由11个连续的α-螺旋区域和8个连续的片层结构区域组成。结合MPBQ-MT蛋白三级结构对MPBQ-MT-基因的生物学功能进一步分析，提出了较为合理的MPBQ-MT作用模型。由系统发生树可知MPBQ-MT蛋白在大豆与菜豆中的亲缘关系较近。
Abstract:
Cloned the 2-methyl-6-phytyl-1,4-benzoquinone methyltransferase (MPBQMTase) of soybean and obtained the cDNA encoding MPBQMTase using Hefeng 25 and Bayfield as materials. The cDNA sequence analysis showed that the cDNA was 1 029 bp in length.Ten latent SNPs were found in CDS of MPBQ-MT gene in soybean by sequences blast.The amino acid sequence and protein structure of MPBQ-MT gene were analyzed by bioinformatics method. The result showed that the CDS region of MPBQ-MT gene encoded 342 amino acid residues with the most content of Valine which accounted for 8.8% of the total amino acids and the least content of Cysteine which accounted for 1.2% of the total amino acids N-terminal of MPBQ-MT protein contains chloroplast transit peptide composed by 58 terminal amino acids and MPBQ-MT was hydrophilic protein. The secondary structure of MPBQ-MT protein was different in Hefeng 25 and Bayfield.About 25.44% and 27.49% of the protein sequence were alpha helixes respectively in the two varieties, about 9.36% and 9.06% of the protein sequence were beta turn, 40.35% and 38.60% of the protein sequence were random coil 24.85% of the protein sequence were lamellar structure in both varieties. S-adenosyl methionine binding sites exist in MPBQ-MT protein of both varieties.The three stage structure analysis showed 11 continuous alpha helixes and 8 continuous film layer structure exist in MPBQ-MT protein. The model of MPBQ-MT was obtained based on the three stage structure of MPBQ-MT protein and biological function analysis of MPBQ-MT. Phylogenetic tree indicated the relationship of MPBQ-MT protein was close in soybean and bean.

参考文献/References:

［1］Sen C K, Khanna S, Roy S. Tocotrienols: Vitamin E beyond tocopherols ［J］. Life Sciences, 2006, 78(18):2088-2098.［2］Fryer M J. The antioxidant effects of thylakoid Vitamin E(α-tocopherol) ［J］. Plant, Cell & Environment, 1992, 15(4): 381-392.

［3］Traber M G, Atkinson J. Vitamin E, antioxidant and nothing more ［J］. Free Radical Biology and Medicine, 2007, 43(1): 4-15.
［4］Vidi P A, Kanwischer M, Baginsky S, et al.Tocopherol cyclase (VTE1) localization and vitamin E accumulation in chloroplast plastoglobule lipoprotein particles ［J］. Journal of Biological Chemistry, 2006, 281(16): 11225-11234.
［5］Della P D, Last R L.Progress in the dissection and manipulation of plant vitamin E biosynthesis ［J］. Physiologia Plantarum, 2006, 126(3): 356-368.
［6］Cheng Z, Sattler S, Maeda H, et al.Highly divergent methyltransferases catalyze a conserved reaction in tocopherol and plastoquinone synthesis in cyanobacteria and photosynthetic eukaryotes ［J］. The Plant Cell, 2003, 15(10): 2343-2356.
［7］李海燕. 大豆维生素E含量的遗传分析及QTL定位 ［D］. 哈尔滨：东北林业大学, 2010:1-10. (Li H Y. Genetic and QTL analysis of the content of vitamin E in soybean ［D］. Northeast Forestry University, 2010:1-10.)
［8］Li H, Liu H, Han Y, et al. Identification of QTL underlying vitamin E contents in soybean seed among multiple environments ［J］. Theoretical and Applied Genetics, 2010, 120(7): 1405-1413.
［9］Gasteiger E, Hoogland C, Gattiker A, et al. Protein identification and analysis tools on the ExPASy server ［M］. Germany:Humana Press, 2005: 571-607.
［10］Emanuelsson O, Nielsen H, Heijne G V. ChloroP, a neural network-based method for predicting chloroplast transit peptides and their cleavage sites ［J］. Protein Science, 1999, 8(5): 978-984.
［11］Wass M N, Kelley L A, Sternberg M J E. 3DLigandSite: Predicting ligand-binding sites using similar structures ［J］. Nucleic Acids Research, 2010: w469-w473.
［12］Hall B G. Building phylogenetic trees from molecular data with MEGA ［J］ Molecular Biology and Evolution, 2013, 30(5): 1229-1235.
［13］Chen Z, Young T E, Ling J, et al. Increasing vitamin C content of plants through enhanced ascorbate recycling ［J］. Proceedings of the National Academy of Sciences, 2003, 100(6): 3525-3530.
［14］Kozbial P Z, Mushegian A R. Natural history of S.adenosylmethionine-binding proteins ［J］. BMC Structural Biology, 2005, 5(1): 19.
［15］朱玉贤, 李毅, 郑晓峰, 等.现代分子生物学［M］4版北京：高等教育出版社, 2013：150-159．(Zhu Y X, Li Y, Zheng X F, et al. Modern molecular biology［M］ 4th ed. Beijing: Higher Education Press, 2013:150-159.)
［16］Vignal A, Milan D, SanCristobal M, et al A review on SNP and other types of molecular markers and their use in animal genetics ［J］. Genetics Selection Evolution, 2002, 34(3): 275-306.
［17］唐立群, 肖层林, 王伟平. SNP分子标记的研究及其应用进展 ［J］. 中国农学通报,2012,12:154-158. (Tang L Q, Xiao C L,Wang W P. Research and application progress of SNP markers ［J］. Chinese Agricultural Science Bulletin, 2012, 12:154-158.)
［18］李殷. 植物维生素E生物合成途径及调控 ［D］. 上海：复旦大学, 2009:2-24. (Li Y.Engineering and regulating plant tocopherol biosynthetic pathway ［D］. Shanghai:Fudan University, 2009:2-24.)
［19］吴光耀. 核编码叶绿体蛋白中的转运肽 ［J］.植物生理学通讯, 1991（3）:166-172.(Wu G Y. Transit peptides for nuclear encoded proteins import into chloroplasts ［J］ Plant Physiology Communications, 1991（3）:166-172.)
［20］Richter S, Lamppa G K. Structural properties of the chloroplast stromal processing peptidase required for its function in transit peptide removal ［J］. Journal of Biological Chemistry, 2003, 278(41): 39497-39502.
［21］Mato J M, Lu S C. Role of S.adenosyl.L.methionine in liver health and injury ［J］ Hepatology, 2007, 45(5): 1306-1312.

相似文献/References:

[1]刘章雄,李卫东,孙石,等.1983～2010年北京大豆育成品种的亲本地理来源及其遗传贡献[J].大豆科学,2013,32(01):1.[doi:10.3969/j.issn.1000-9841.2013.01.002]
　LIU Zhang-xiong,LI Wei-dong,SUN Shi,et al.Geographical Sources of Germplasm and Their Nuclear Contribution to Soybean Cultivars Released during 1983 to 2010 in Beijing[J].Soybean Science,2013,32(01):1.[doi:10.3969/j.issn.1000-9841.2013.01.002]
[2]李彩云,余永亮,杨红旗,等.大豆脂质转运蛋白基因GmLTP3的特征分析[J].大豆科学,2013,32(01):8.[doi:10.3969/j.issn.1000-9841.2013.01.003]
　LI Cai-yun,YU Yong-liang,YANG Hong-qi,et al.Characteristics of a Lipid-transfer Protein Gene GmLTP3 in Glycine max[J].Soybean Science,2013,32(01):8.[doi:10.3969/j.issn.1000-9841.2013.01.003]
[3]王明霞,崔晓霞,薛晨晨,等.大豆耐盐基因GmHAL3a的克隆及RNAi载体的构建[J].大豆科学,2013,32(01):12.[doi:10.3969/j.issn.1000-9841.2013.01.004]
　WANG Ming-xia,CUI Xiao-xia,XUE Chen-chen,et al.Cloning of Halotolerance 3 Gene and Construction of Its RNAi Vector in Soybean (Glycine max)[J].Soybean Science,2013,32(01):12.[doi:10.3969/j.issn.1000-9841.2013.01.004]
[4]张春宝,李玉秋,彭宝,等.线粒体ISSR与SCAR标记鉴定大豆细胞质雄性不育系与保持系[J].大豆科学,2013,32(01):19.[doi:10.3969/j.issn.1000-9841.2013.01.005]
　ZHANG Chun-bao,LI Yu-qiu,PENG Bao,et al.Identification of Soybean Cytoplasmic Male Sterile Line and Maintainer Line with Mitochondrial ISSR and SCAR Markers[J].Soybean Science,2013,32(01):19.[doi:10.3969/j.issn.1000-9841.2013.01.005]
[5]卢清瑶,赵琳,李冬梅,等.RAV基因对拟南芥和大豆不定芽再生的影响[J].大豆科学,2013,32(01):23.[doi:10.3969/j.issn.1000-9841.2013.01.006]
　LU Qing-yao,ZHAO Lin,LI Dong-mei,et al.Effects of RAV gene on Shoot Regeneration of Arabidopsis and Soybean[J].Soybean Science,2013,32(01):23.[doi:10.3969/j.issn.1000-9841.2013.01.006]
[6]杜景红,刘丽君.大豆fad3c基因沉默载体的构建[J].大豆科学,2013,32(01):28.[doi:10.3969/j.issn.1000-9841.2013.01.007]
　DU Jing-hong,LIU Li-jun.Construction of fad3c Gene Silencing Vector in Soybean[J].Soybean Science,2013,32(01):28.[doi:10.3969/j.issn.1000-9841.2013.01.007]
[7]张力伟,樊颖伦,牛腾飞,等.大豆“冀黄13”突变体筛选及突变体库的建立[J].大豆科学,2013,32(01):33.[doi:10.3969/j.issn.1000-9841.2013.01.008]
　ZHANG Li-wei,FAN Ying-lun,NIU Teng-fei?,et al.Screening of Mutants and Construction of Mutant Population for Soybean Cultivar "Jihuang13”[J].Soybean Science,2013,32(01):33.[doi:10.3969/j.issn.1000-9841.2013.01.008]
[8]盖江南,张彬彬,吴瑶,等.大豆不定胚悬浮培养基因型筛选及基因枪遗传转化的研究[J].大豆科学,2013,32(01):38.[doi:10.3969/j.issn.1000-9841.2013.01.009]
　GAI Jiang-nan,ZHANG Bin-bin,WU Yao,et al.Screening of Soybean Genotypes Suitable for Suspension Culture with Adventitious Embryos and Genetic Transformation by Particle Bombardment[J].Soybean Science,2013,32(01):38.[doi:10.3969/j.issn.1000-9841.2013.01.009]
[9]王鹏飞,刘丽君,唐晓飞,等.适于体细胞胚发生的大豆基因型筛选[J].大豆科学,2013,32(01):43.[doi:10.3969/j.issn.1000-9841.2013.01.010]
　WANG Peng-fei,LIU Li-jun,TANG Xiao-fei,et al.Screening of Soybean Genotypes Suitable for Somatic Embryogenesis[J].Soybean Science,2013,32(01):43.[doi:10.3969/j.issn.1000-9841.2013.01.010]
[10]刘德兴,年海,杨存义,等.耐酸铝大豆品种资源的筛选与鉴定[J].大豆科学,2013,32(01):46.[doi:10.3969/j.issn.1000-9841.2013.01.011]
　LIU De-xing,NIAN Hai,YANG Cun-yi,et al.Screening and Identifying Soybean Germplasm Tolerant to Acid Aluminum[J].Soybean Science,2013,32(01):46.[doi:10.3969/j.issn.1000-9841.2013.01.011]
[11]刘焕成,李文滨,韩英鹏,等.大豆维生素E与主要农艺性状和品质性状的相关性分析[J].大豆科学,2011,30(01):89.[doi:10.11861/j.issn.1000-9841.2011.01.0089]
　LIU Huan-cheng,LI Wen-bin,HAN Ying-peng,et al.Correlation Analysis among Vitamin E Content, Agronomic Traits and Quality Characteristics of Soybean[J].Soybean Science,2011,30(01):89.[doi:10.11861/j.issn.1000-9841.2011.01.0089]
[12]李海燕,韩英鹏,武小霞,等.大豆维生素E遗传图谱构建及QTL分析[J].大豆科学,2014,33(04):492.[doi:10.11861/j.issn.1000-9841.2014.04.0492]
　LI Hai-yan,HAN Ying-peng,WU Xiao-xia,et al.QTL Analysis of Soybean Vitamin E and Genetic Map Construction[J].Soybean Science,2014,33(01):492.[doi:10.11861/j.issn.1000-9841.2014.04.0492]
[13]刘焕成,韩英鹏,腾卫丽,等.东北大豆与北美大豆维生素E含量的分析[J].大豆科学,2008,27(06):925.[doi:10.11861/j.issn.1000-9841.2008.06.0925]
　LIU Huan-cheng,HAN Ying-peng,TENG Wei-li,et al.Analysis of Vitamin E Content in Soybeans Derived from Northeast China and North America[J].Soybean Science,2008,27(01):925.[doi:10.11861/j.issn.1000-9841.2008.06.0925]
[14]罗健,冯雷,李冬梅,等.不同大豆品种籽粒维生素E含量积累比较分析[J].大豆科学,2017,36(02):250.[doi:10.11861/j.issn.1000-98412017020250]
　LUO Jian,FENG Lei,LI Dongmei,et al.Comparative Analysis of Vitamin E Content Accumulation in Different Soybean Varieties[J].Soybean Science,2017,36(01):250.[doi:10.11861/j.issn.1000-98412017020250]
[15]刘焕成,曹广禄,鹿娜,等.大豆维生素E含量与农艺性状及品质性状的相关性分析[J].大豆科学,2018,37(02):224.[doi:10.11861/j.issn.1000-9841.2018.02.0224]
　LIU Huan-cheng,CAO Guang-lu,LU Na,et al.Correlation Analysis Among Vitamin E Content, Agronomic Traits, Protein and Oil Contents of Soybean[J].Soybean Science,2018,37(01):224.[doi:10.11861/j.issn.1000-9841.2018.02.0224]

备注/Memo

基金项目：中国博士后项目（20110491025）；东北农业大学博士启动基金（2012RCB32）。

第一作者简介：聂腾坤（1990-），男，硕士，主要从事大豆遗传育种研究。E-mail:Chinantk@126.com。
通讯作者：李海燕（1978-），女，博士，副研究员，主要从事大豆遗传育种及生物技术研究。E-mail: lhy2112002@163.com；李文滨（1958-），男，教授，博导，主要从事大豆遗传育种研究。E-mail：wenbinli@neau.edu.cn。

更新日期/Last Update: 2016-01-28