Cloning and Expression Analysis of Actin ( MwACT 2) in Agropyron mongolicum Keng

doi:10.11733/j.issn.1007-0435.2016.01.018

Acta Agrestia Sinica ›› 2016, Vol. 24 ›› Issue (1): 129-136.DOI: 10.11733/j.issn.1007-0435.2016.01.018

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Cloning and Expression Analysis of Actin ( MwACT 2) in Agropyron mongolicum Keng

AO Te-gen-bai-yin^1,5, GAO Li-jie⁵, ZHANG Shu-hua², YUN Jing-feng³, LANG Ming-lin², YANG Xue-ju^2,4

1. College of Agronomy, Agricultural University of Hebei, Baoding, Hebei Province 071000, China;
2. College of Life Sciences, Agricultural University of Hebei, Baoding, Hebei Province 071000, China;
3. College of Ecology and Environment, Inner Mongolia Agricultural University, Huhhot, Inner Mongolia 010019, China;
4. Key Laboratory of Crop Germplasm in Hebei Province, Baoding, Hebei Province 071000, China;
5. College of Animal Science and Technology,Agriculture University of Hebei, Baoding, Hebei Province 071000, China

Received:2014-12-23 Revised:2015-10-17 Online:2016-02-15 Published:2016-04-26

蒙古冰草肌动蛋白基因(MwACT2)克隆与表达分析

敖特根白音^1,5, 高立杰⁵, 张树华², 云锦凤³, 郎明林², 杨学举^2,4

1. 河北农业大学农学院, 河北保定 071001;
2. 河北农业大学生命科学院, 河北保定 071001;
3. 内蒙古农业大学生态环境学院, 内蒙古呼和浩特 010018;
4. 河北省作物种质资源实验室, 河北保定 071001;
5. 河北农业大学动物科技学院, 河北保定 071001

通讯作者: 郎明林, 杨学举
作者简介:敖特根白音(1975-),男,蒙古族,内蒙古赤峰人,博士,讲师,从事牧草开发育种研究,E-mail:atg12@sina.com
基金资助:
河北省科技支撑项目(06220114D);河北省现代农业产业技术体系建设项目(1004002);河北省百名优秀创新人才支持计划(0901083);河北生应用基础研究计划重点基础项目(14966318D)资助

Abstract

Abstract:

An Actin gene of Agropyron mongolicum Keng was isolated by RT-PCR and RACE, was named as MwACT2. Sequence analysis indicated that the full-length of MwACT2 gene is 1506 bp and contained an 1131 bp open reading frame. It encodes a polypeptide of 376 amino acids. The estimated molecular weightof the putative MwACT2 protein was 41.606 kDa with an isoelectric point of 5.29 . MwACT2 has a highly conserved actin domain, which demonstrated it is a member of actin family. MwACT2 was predicted to be a non-transmembrane protein. The phylogenetic relationship also indicated that MwACT2 protein was similar to Triticum urartu (accession no. EMS62134.1) with amino acid sequences similarity of 99%. Result of semi-quantitative RT-PCR showed that the expression of MwACT2 was constant. There was no significant difference under high salt, drought and cold treatment.

Key words: Agropyron mongolicum Keng, Actin gene, Gene cloning, 5' RACE, Bioinformatics analysis, Expression analysis

摘要：

本研究应用RT-PCR和RACE技术,从蒙古冰草(Agropyron mongolicum Keng)中分离到1个肌动蛋白基因,命名为MwACT2。序列分析表明:MwACT2基因全长1506bp,含有1个1131bp的开放型阅读框,MwACT2蛋白长度为376个氨基酸残基,分子量为41.606KDa,理论等电点为5.29,含有actin保守位点,属于actin家族成员。该基因编码的氨基酸序列与Genebank中乌拉尔图(Triticum_urartu)肌动蛋白(登录号:EMS62134.1)氨基酸具有99%的一致性,属于非跨膜蛋白。利用半定量RT-PCR技术分析MwACT2基因的表达,结果表明该基因表达稳定,经高盐、干旱、低温处理后的表达强度无显著差异。

关键词: 蒙古冰草, 肌动蛋白基因, 基因克隆, 5'RACE, 生物信息学分析, 表达分析

CLC Number:

AO Te-gen-bai-yin, GAO Li-jie, ZHANG Shu-hua, YUN Jing-feng, LANG Ming-lin, YANG Xue-ju. Cloning and Expression Analysis of Actin ( MwACT 2) in Agropyron mongolicum Keng[J]. Acta Agrestia Sinica, 2016, 24(1): 129-136.

敖特根白音, 高立杰, 张树华, 云锦凤, 郎明林, 杨学举. 蒙古冰草肌动蛋白基因(MwACT2)克隆与表达分析[J]. 草地学报, 2016, 24(1): 129-136.

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Cloning and Expression Analysis of Actin ( MwACT 2) in Agropyron mongolicum Keng

蒙古冰草肌动蛋白基因(MwACT2)克隆与表达分析

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