甘蔗锌指蛋白基因ShSAP1的克隆与表达模式

Cloning and expression pattern of a zinc finger protein gene ShSAP1 in Saccharum officinarum

植物中具有A20/AN1锌指结构域的蛋白与逆境应答密切相关,在甘蔗热带种Saccharum officinarum拔地拉Badila中克隆到一个具有A20/AN1锌指结构域的锌指蛋白基因ShSAP1。为研究ShSAP1的基因结构和表达特性,采用PCR和Southern blotting分析了ShSAP1的基因组结构,通过半定量RT-PCR对ShSAP1在甘蔗不同部位、不同胁迫和不同激素处理下的表达模式进行了分析。结果表明,ShSAP1的5＇UTR区有两段内含子,大小分别为202 bp和1 052 bp,在Badila中有1~2个拷贝;ShSAP1在甘蔗成熟植株根茎叶中均有表达,在茎秆中的表达量随着茎节成熟而递增;在幼苗阶段,ShSAP1的表达能被高盐、模拟干旱、GA3、ABA和ET诱导增强。以上结果表明,ShSAP1可能在甘蔗成熟与逆境应答中具有重要作用。

In plants,proteins with A20/AN1 zinc finger domain are involved in stress responses,named as ＂Stress Associated Protein＂（SAP） gene family.Based on Expressed Sequence Tag（EST） sequences information in Badila Saccharum officinarum mature related cDNA library,we cloned an SAP gene from sugarcane full length cDNA library,named ShSAP1（GenBank： Accession No.HM991960）.To characterize ShSAP1,we analyzed its genome structure and expression pattern.Southern blot analysis showed ShSAP1 was present as one or two copy in the genome of Badila.Comparison of ShSAP1 1 008 bp full length cDNA with a genomic frangment（2 241 bp） generated by PCR amplification and sequencing,revealed the presence of two introns（202 bp and 1 052 bp） located in the 5＇UTR region.Semiquantitative RT-PCR analysis found ShSAP1 expressed in leaves,roots and stalk in mature sugarcane.Compared with immature stems,ShSAP1 expressed higher in mature stalk.ShSAP1 was induced by different types of treatments,such as salt（200 mmol/L NaCl）,drought（10% PEG 6 000）,GA3（200 mg/L）,ABA（100 μmol/L） and ET（1 mmol/L） during sugarcane seedling stage.These results indicated that ShSAP1 may function in sugarcane maturation and abiotic stress response processes.