<em>Ipomoeatrifida</em>(2X)抗坏血酸过氧化物酶基因的克隆、序列分析和表达分析

doi:10.7668/hbnxb.2015.02.003

华北农学报 ›› 2015, Vol. 30 ›› Issue (2): 12-16. doi: 10.7668/hbnxb.2015.02.003

所属专题： 生物技术；

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

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

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

收稿日期:2015-02-23 出版日期:2015-04-28
通讯作者: 唐君(1960-),女,新疆奎屯人,研究员,主要从事甘薯资源评价及组培技术研究。
作者简介:张安(1984-),男,山东菏泽人,助理研究员,博士,主要从事甘薯野生资源利用研究。
基金资助:
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)

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

ZHANG An^1,2, CAO Qing-he^1,2, ZHOU Zhi-lin^1,2, ZHAO Dong-lan^1,2, PANG Hong-bo³, TANG Jun^1,2

1. Key Laboratory of Biology and Genetic Breeding of Sweet Potato, Ministry of Agriculture, Xuzhou 221131, China;
2. Xuhuai District Institute of Agricultural Sciences, Xuzhou 221131, China;
3. Chemistry and Life Science College, Shenyang Normal University, Shenyang 110034, China

Received:2015-02-23 Published:2015-04-28

摘要/Abstract

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cDNA,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 kDa,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,ItAPX和IbAPX最为接近;组织特异性表达分析表明, ItAPX 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。

关键词: 甘薯, Ipomoea trifida, APX, 抗氧化

Abstract: In order to get stress resistance genes from wild resources of sweet potato, total RNA was extracted from tissue culture seedlings of Ipomoea 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 protein 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 expressed in all organs.Cloning of I.trifida ascorbate peroxidase gene would help sweet potato breeding for stress-tolerance with new genetic resources.

Key words: Sweet potato, Ipomoea trifida, APX, Anti-oxidation

中图分类号:

S531.03
Q78

张安, 曹清河, 周志林, 赵冬兰, 逄洪波, 唐君. Ipomoeatrifida(2X)抗坏血酸过氧化物酶基因的克隆、序列分析和表达分析[J]. 华北农学报, 2015, 30(2): 12-16. doi: 10.7668/hbnxb.2015.02.003.

ZHANG An, CAO Qing-he, ZHOU Zhi-lin, ZHAO Dong-lan, PANG Hong-bo, TANG Jun. Cloning,Bioinformatic Analysis and Expression Analysis of APX Gene from Ipomoea trifida (2X)[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2015, 30(2): 12-16. doi: 10.7668/hbnxb.2015.02.003.

http://www.hbnxb.net/CN/Y2015/V30/I2/12

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Ipomoeatrifida(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)

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