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锌指蛋白是一类具有手指型结构的蛋白质,其中一些锌指蛋白是转录因子,对真核生物的生长发育及非生物逆境胁迫的耐受能力都有着重要作用。文章从大豆(Glycine max(L.)Merr.)中克隆了一个新的C2H2型锌指蛋白基因SCTF-1(GenBank登录号:JQ692081),该基因包含一个699 bp的开放阅读框,编码233个氨基酸,无内含子,有两个典型的C2H2型锌指结构。锌指结构中有植物锌指蛋白特有的保守氨基酸序列QALGGH。经软件预测分析,其等电点pI=8.33,分子量24.9 kDa。农杆菌介导的洋葱表皮细胞GFP瞬时表达实验结果表明,SCTF-1蛋白能够定位到细胞核中。通过RT-PCR检测发现该基因在大豆叶和花中的表达量较高,在茎和根的表达量相对较低。在对大豆幼苗的低温胁迫中,SCTF-1基因的表达量明显增加。将SCTF-1基因转入烟草(Nicotiana tabacum L.)中,发现SCTF-1基因的过量表达能够明显提高转基因烟草的耐冷能力。
Zinc finger proteins are a class of finger-type proteins, some of which are transcription factors and play an important role in the growth and development of eukaryotes and the tolerance of abiotic stresses. A novel C2H2-type zinc finger protein gene SCTF-1 (GenBank accession number: JQ692081) was cloned from Glycine max (L.) Merr. The gene contains a 699 bp open reading frame encoding 233 Amino acids, without introns, have two typical C2H2-type zinc finger structures. Zinc finger structure zinc plant zinc finger protein unique conserved amino acid sequence QALGGH. Predicted by software, its isoelectric point pI = 8.33, molecular weight 24.9 kDa. Agrobacterium-mediated transient expression of GFP in onion epidermal cells showed that SCTF-1 protein could be localized to the nucleus. The result of RT-PCR showed that the gene was highly expressed in soybean leaf and flower and relatively low in stem and root. In the cold stress on soybean seedlings, the expression of SCTF-1 gene was significantly increased. The SCTF-1 gene was transferred into tobacco (Nicotiana tabacum L.), and it was found that overexpression of SCTF-1 gene can significantly increase the cold tolerance of transgenic tobacco.