摘要：

锌指蛋白是真核生物中被广泛研究的转录因子,在植物生长发育和逆境应答中发挥重要作用。为了揭示大豆锌指蛋白基因功能,从商豆1201中克隆获得GmZAT12基因的CDS全长序列,并对其编码的蛋白质进行生物信息学分析。通过烟草表皮注射系统检测GmZAT12蛋白的亚细胞定位情况,采用实时荧光定量(qRT-PCR)技术对GmZAT12基因在大豆不同组织和非生物胁迫中的表达模式进行分析。结果表明,GmZAT12全长516 bp,共编码171个氨基酸,分子质量为19.264 28 ku,理论等电点(pI)为9.02;主要构成元件为无规则卷曲和α螺旋;含有20个磷酸化位点,其中以丝氨酸磷酸化位点为主。序列分析结果表明,GmZAT12蛋白含有2个保守的C2H2锌指结构域;亚细胞定位结果显示,GmZAT12蛋白定位于细胞核。qRT-PCR表达分析结果显示,GmZAT12基因在大豆根、叶片和种子中表达量较高,在花和茎中表达量较低;GmZAT12基因受到高温、低温、NaCl和ABA诱导表达,推测该基因可能参与大豆非生物胁迫应答。

关键词: 大豆, GmZAT12基因, 克隆, 表达分析, qRT-PCR

Abstract:

Zinc finger proteins are transcription factors widely studied in eukaryotes,and play important roles in plant growth and development,and responses to stresses.In order to deeply understand the gene function of zinc finger protein in soybean,the full-length CDS sequence of GmZAT12 gene was cloned from Shangdou 1201,and the characteristics of coded protein by this gene was predicted by bioinformatics analysis.The subcellular localization of GmZAT12 protein was detected by the tobacco epidermal injection system, and the expression pattern of GmZAT12 gene in different tissues of soybean and abiotic stress was analyzed by Real-time PCR (qRT-PCR) technology. The results showed that GmZAT12 CDS contained 516 bp,encoding 171 amino acids and the molecular weight of the protein was 19.264 28 ku with a theoretical isoelectric point(pI)of 9.02.Its main components were random coils and α-helices and it contained 20 phosphorylation sites, mainly serine phosphorylation sites.Sequence analysis indicated that GmZAT12 possessed two conserved C2H2 zinc finger domains.The result of subcellular location indicated that GmZAT12 protein was localized in the nucleus.The results of qRT-PCR showed that CmZAT12 gene expressed mainly in roots,leaves and seeds of soybean,while low expression in flower and stem, and was induced by high temperature,low temperature,NaCl and ABA.The fact implied that this gene might be involved in abiotic stress signaling pathways.

Key words: Soybean, GmZAT12 gene, Cloning, Expression analysis, qRT-PCR