| 摘要: |
| 以大豆叶片总RNA为模板,采用RT-PCR法从吉豆2号品种中克隆获得大豆种子成熟蛋白(PM34)基因序列,利用生物信息学方法对大豆PM34基因编码的蛋白进行预测。结果表明:该基因编码蛋白理论分子量为31.7 kDa,等电点为6.60,为亲水性蛋白; 该蛋白中无跨膜结构; 该蛋白中不存在信号肽。PM34基因编码蛋白的二级结构中α 螺旋占12.97%,无规则卷曲占41.30%,β折叠占45.73%。PM34基因编码蛋白的三级结构预测表明,同源模建的模板是3ijr.1.A,是一种短链脱氢酶/还原酶,与该蛋白的同源性为54.65%。在进化关系上,与绿豆、苜蓿的亲缘关系相对较近。采用实时定量PCR方法(qRT-PCR),检测大豆PM34基因在大豆各器官中的表达方式,结果表明该基因在大豆根、茎、叶、花中的表达活性低,而在种子中,尤其是成熟种子中的相对表达活性很高。该研究结果为大豆PM34基因结构和功能的进一步研究奠定了基础。 |
| 关键词: 大豆, PM34基因, 克隆, 生物信息学, qRT-PCR |
| DOI：10.11931/guihaia.gxzw201601026 |
| 分类号:Q943.2 |
| 文章编号:1000-3142(2016)10-1220-05 |
| Fund project:黑龙江省自然科学基金(C201458); 黑龙江省普通本科高等学校青年创新人才培养计划(UNPYSCT_2016090); 国家自然科学基金(31301335)[Supported by the Natural Science Foundation of Heilongjiang Province(C201458); Training Plan of Young Creative Talents of Average Four-Year College of Heilongjiang(UNPYSCT_2016090); the National Nature Science Foundation of China(31301335)]。 |
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| Bioinformatics and the expression pattern analysis of soybean PM34 gene |
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ZHAO Yan1*, ZHAI Ying1, GONG Guo-Qiang2,
WANG Man1, ZHAO Yang1
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1. College of Life Sciences and Agroforestry, Qiqihaer University, Qiqihaer 161006, Heilongjiang, China;2. Jilin Provincial Institute of Food Inspection, Changchun 130000, China
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| Abstract: |
| In this paper, the structure and tissue-specificity of the soybean seeds mature protein gene( PM34 )were studied, which could become the the foundations for further research the function of PM34 gene in soybean or the other plants. The sequence of soybean(Jidou2)PM34 gene was amplified by RT-PCR with the total RNA of soybean leaf as templet. The structure of soybean PM34 gene was predicted by bioinformatics. The expression pattern of soybean PM34 gene in soybean organs was detected by real-time quantitative PCR(qRT-PCR). The results showed that the molecular weight of the protein was 31.7 kDa, the isoelectric point was 6.60, and belonging to the hydrophilic protein. There was absent transmembrane domain and singal peptide. The secondary structure was comprosed of 12.97% alpha helix, 41.30% coil and 45.73% extended strand. The protein tertiary structure prediction showed that the homologous model template was 3 ijr. 1. A, which was a kind of short chain dehydrogenase/reductase and the homology was 54%. The evolutionary relationship demonstrated that soybean had relatively closer relationship with Vigna radiata and Medicago trun-catula than others. In addtion, the result showed that there was a little activity in roots, stems, leaves and flowers, but there was the highest activity in seeds, especially in mature seeds. |
| Key words: soybean, PM34 gene, cloning, bioinformaics, qRT-PCR |
