摘要ERF是一种带有ERF结构域的转录因子，是一种与植物抗逆境相关的调控因子，可以帮助植物抵抗低温、高盐、干旱等非生物胁迫。根据已报道的番茄ERF转录因子家族的基因表达信息，筛选得到一个受盐和干旱胁迫显著诱导的SlERF16基因，并根据其基因序列设计特异引物，采取RT-PCR（反转录PCR）的方法在野生型番茄AC++中克隆出该基因的cDNA全长序列。序列分析表明，SlERF16基因开放阅读框全长为747 bp，编码了248个氨基酸。生物信息学分析表明，其编码的氨基酸具有ERF蛋白家族典型的AP2结构域。蛋白分子量为27.84 kD，理论等电点pI为4.61，疏水性平均值为-0.647，说明该蛋白为亲水性蛋白。磷酸化位点预测显示该蛋白具有多个磷酸化位点，亚细胞定位分析表明其定位于细胞核。多重序列比对分析显示，SlERF16蛋白与其他植物ERF蛋白具有很高的同源性。这些结果可以为更进一步的研究SlERF16基因在番茄抗逆境响应中的作用提供理论依据。71130

该论文有图11幅，表1个，参考文献24篇。

毕业论文关键词：番茄;  ERF转录因子； SlERF16； 生物信息学分析

Cloning and bioinformatics analysis of stress response SlERF16 gene in Tomato

Abstract ERF is a transcription factor with ERF domain structure. It is a regulatory factor related to plant resistance. They can help the plant improve the resistance to low temperature, drought and other abiotic stresses. According to the reported gene expression information of tomato ERF transcription factors, we got the SlERF16 gene induced significantly by salt and drought stress. Specific primer was designed to clone the full length cDNA sequence of SlERF16 gene from wild type tomato AC++. Sequence analysis showed that the open reading frame of SlERF16 gene is 747 bp, encoding 248 amino acids. Bioinformatics analysis showed that the encoded amino acid has a typical AP2 domain structure of the ERF protein family. The molecular weight of SlERF16 protein is 27.84 kD, isoelectric point pI is 4.61, and its GRAVY is -0.647, indicating that the protein is hydrophilic protein. Phosphorylation sites prediction showed that the protein has multiple phosphorylation sites, subcellular localization analysis showed that its localization is the nucleus. Multiple sequence alignment analysis showed that SlERF16 protein has high homology with the ERF proteins from other plants. These results laid a foundation for the further study of SlERF16’s function in response to stress of tomato.

Key  words: tomato;   ERF Transcription factor;   SlERF16;   bioinformatics   analysis            

目录

摘要 I

Abstract II

目录 III

图清单 IV

表清单 IV

1绪论 5

2 材料与试剂 6

2.1 材料 6

2.2试剂 6

3 仪器与方法 6

3.1 仪器 7

3.2 方法 7

4实验结果与分析 10

4.1 番茄总RNA的提取 11

4.2 SlERF16基因特异性引物的设计及合成 11

4.3 基因的PCR扩增 12

4.4 菌落PCR 12