Ipomoea trifida(2X)抗坏血酸过氧化物酶基因的克隆、序列分析和表达分析  

Cloning,Bioinformatic Analysis and Expression Analysis of APX Gene from Ipomoea trifida( 2X)

作　　者：张安[1,2] 曹清河[1,2] 周志林[1,2] 赵冬兰[1,2] 逄洪波[3] 唐君[1,2] 

机构地区：[1]农业部甘薯生物学与遗传育种重点实验室,江苏徐州221131 [2]江苏徐淮地区徐州农业科学研究所,江苏徐州221131 [3]沈阳师范大学化学与生命科学学院,辽宁沈阳110034

出　　处：《华北农学报》2015年第2期12-16,共5页Acta Agriculturae Boreali-Sinica

基　　金：国家甘薯产业技术体系项目(CARS-11-B-02-2014);国家自然科学基金项目(31100176);江苏省自然科学基金面上项目(BK20131125)

摘　　要：为了从甘薯野生资源中获取抗逆基因,提取Ipomoea trifida组培苗总RNA,经反转录获得第一链c DNA,使用特异引物进行PCR扩增,获得了I.trifida抗坏血酸过氧化物酶编码区序列,并进行生物信息学分析以及组织特异性表达研究。经测序表明,所获得的序列长约769 bp,包含753 bp完整的开放阅读框;生物信息学分析结果表明,此序列编码250个氨基酸,蛋白大小约为27.6 k Da,等电点为5.42,有大范围亲水区,推测在细胞质中起到抗氧化作用;进化发育系统分析表明,It APX和Ib APX最为接近;组织特异性表达分析表明,It APX在各组织中均有表达。I.trifida抗坏血酸过氧化物酶基因的克隆为甘薯抗逆育种提供了新的基因资源。In order to get stress resistance genes from wild resources of sweet potato,total RNA was extracted from tissue culture seedlings of Iponoea trifida,cDNA first-strand was obtained by reverse transcription,a 769 bp sequence was amplified by PCR using specific primers. The I. trifida ascorbate peroxidase enzyme coding region was obtained,bioinformatic analysis and expression pattern analysis were conducted. Sequencing results showed that the sequence contained a complete open reading frame of 753 bp encoding 250 amino acid residues. The putative pro-tein size was approximately 27. 6 kDa,isoelectric point was 5. 42,and there was a wide range of hydrophilic region. It was speculated that the protein played antioxidant role in the cytoplasm. Phylogenetic analysis indicated that IbAPX was most closely genetically related to ItAPX. Tissue-specific expression analysis showed that ItAPX ex-pressed in all organs. Cloning of I. trifida ascorbate peroxidase gene would help sweet potato breeding for stress-tol-erance with new genetic resources.

关 键 词：甘薯 IPOMOEA trifida APX 抗氧化 

分 类 号：S531.03[农业科学—作物学] Q78[生物学—分子生物学]