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应用RT-PCR技术,从玉米自交系承18经14℃冷锻炼处理的芽鞘中扩增出长1491 bp的Cat3全长cDNA。以pGEM-T Easy为载体,克隆了扩增片段,经酶切鉴定和DNA序列测定,确认为玉米Cat3基因。序列分析结果表明,该序列与GenBank中accession number:L05934的序列仅在两个核苷酸位点存在差异。同时采用Northern杂交方法对Cat3在不同低温处理的玉米芽鞘中的表达特性进行了研究,结果显示Cat3在经14℃冷锻炼处理的芽鞘中表达增强,表明该基因受低温的诱导表达,为玉米抗寒基因,对提高玉米的抗寒力起积极作用。
The full-length cDNA of 1491 bp was amplified by RT-PCR from the sheath of maize inbred line 18 at 14 ℃ cold treatment. Using pGEM-T Easy as vector, the amplified fragment was cloned and identified as the maize Cat3 gene by enzyme digestion and DNA sequencing. Sequence analysis showed that there was a difference between the sequence and accession number: L05934 in GenBank at only two nucleotide positions. At the same time, Northern hybridization method was used to study the expression characteristics of Cat3 in different low-temperature treatment of corn germ sheath. The results showed that Cat3 expression in the sheath of the sheath after cold acclimation treatment at 14 ℃, indicating that the gene induced by low temperature, Cold-resistant maize genes play a positive role in improving the cold resistance of maize.