柽柳过氧化物酶基因的序列分析及山新杨遗传转化研究

Sequence Analysis of Peroxidase Gene from Tamarix hispida and Its Transformation Into Populus davidiana×P.bolleana

对刚毛柽柳(Tamarix hispida)的过氧化物酶基因(ThPOD)全长序列进行分析,结果表明,该基因全长为1 376 bp,包含一个1 086 bp的开放阅读框,可编码361个氨基酸残基,蛋白分子质量为39.3 ku。根据ThPOD基因保守序列设计特异引物克隆该基因,并成功构建了pROKⅡ-ThPOD植物过表达载体,利用农杆菌介导法转化山新杨(Populus davidiana×P.bolleana),经PCR和Northern blot鉴定,初步证明外源基因ThPOD已整合到山新杨的基因组中并在转录水平上进行了表达。

The sequence of ThPOD gene isolated from Tamarix hispida root cDNA library was analyzed and it was showed that the sequence length of ThPOD was 1 376 kb containing a 1 086 bp ORF.The protein molecular quality of ThPOD was 39.3 ku with 361 amino acids.The gene was cloned from the specific primers designed according to the conserved sequences of ThPOD.Plant over-expression vector pROKⅡ-ThPOD was constructed;And the ThPOD gene was transformed into Populus davidiana × P.bolleana through Agrobacterium-mediated genetic transformation.PCR and Northern blot results showed that ThPOD was integrated into the genome and the transformed gene was expressed at the transcription level.