[1]张丽丽,徐碧玉,刘菊华,等.香蕉MaASR1基因的生物信息学分析及原核表达[J].江苏农业学报,2017,(05):1129-1135.[doi:doi:10.3969/j.issn.1000-4440.2017.05.026]
　ZHANG Li-li,XU Bi-yu,LIU Ju-hua,et al.Bioinformatics analysis and prokaryotic expression of MaASR1 gene from banana[J].,2017,(05):1129-1135.[doi:doi:10.3969/j.issn.1000-4440.2017.05.026]
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香蕉MaASR1基因的生物信息学分析及原核表达()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2017年05期
页码:
1129-1135
栏目:
园艺
出版日期:
2017-10-30

文章信息/Info

Title:
Bioinformatics analysis and prokaryotic expression of MaASR1 gene from banana
作者:
张丽丽1; 徐碧玉1; 刘菊华1; 贾彩红1; 张建斌1; 金志强1; 2
（1.中国热带农业科学院热带生物技术研究所/农业部热带作物生物学与遗传资源利用重点实验室，海南海口571101；2.中国热带农业科学院海口实验站/海南省香蕉遗传育种改良重点实验室，海南海口570102）
Author(s):
ZHANG Li-li1; XU Bi-yu1; LIU Ju-hua1; JIA Cai-hong1; ZHANG Jian-bin1; JIN Zhi-qiang1; 2
(1.Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Sciences/Key Laboratory of Biology and Genetic Resources of Tropical Crops, Ministry of Agriculture, Haikou 571101, China;2.Haikou Experimental Station, Chinese Academy of Tropical Agricultural Sciences/Hainan Provincial Key Laboratory for Genetics and Breeding of Banana, Haikou 570102, China)
关键词:
MaASR1基因; 原核表达; 生物信息学; 蛋白质纯化
Keywords:
MaASR1 gene; prokaryotic expression; bioinformatics; protein purification
分类号:
S668.1
DOI:
doi:10.3969/j.issn.1000-4440.2017.05.026
文献标志码:
A
摘要:
为了深入研究MaASR1基因的分子生物学功能，利用RT-PCR技术从拟南芥转基因株系L14中克隆到MaASR1基因的 cDNA 序列，该序列含有1个432 bp 的开放阅读框，编码143个氨基酸的蛋白质，并对该序列进行了生物信息学分析。将该基因克隆到原核表达载体PET-30a中，经酶切和测序鉴定后，将正确的重组质粒pET30a-MaASR1导入大肠杆菌BL21(DE3)，成功构建了该基因的原核表达载体。利用所构建的原核表达载体进行原核表达，经IPTG诱导和SDS-PAGE电泳检测，结果表明表达蛋白质与预期蛋白质大小基本一致。利用Ni柱亲和层析方法进一步获得了纯度较高的重组蛋白质，并用Western blot方法确定此重组蛋白质为目的蛋白质。
Abstract:
In order to study the molecular biological function of MaASR1 gene, the cDNA sequence of MaASR1 from Arabidopsis thaliana transgenic line L14 was cloned by RT-PCR. The open reading frame of MaASR1 was 432 bp in length，which encoded 143 amino acids sequence. The sequence was analyzed by bioinformatics. MaASR1 gene was inserted into the prokaryotic expression vector PET-30a. After confirmation by digestion and sequencing，the recombinant plasmid pET30a-MaASR1 was transformed into Escherichia coli strain BL21( DE3). The recombinant protein with the predicted molecular weight was successfully induced to express using IPTG and was detected and confirmed by SDS-PAGE and Western blot. The highly purified recombinant protein was obtained by Ni affinity chromatography.

参考文献/References:

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备注/Memo

备注/Memo:
收稿日期：2017-07-03 基金项目：转基因生物新品种培育国家科技重大专项（2016ZX08012005-007）；国家香蕉产业技术体系品种改良项目（CARS-31）；中央级公益性科研院所基本科研业务费专项（1630052017012） 作者简介：张丽丽（1984-），女，山东滨州人，博士，助理研究员，主要从事植物分子遗传学研究。（Tel）0898-66894828；（E-mail）zhanglili@itbb.org.cn 通讯作者：金志强，（E-mail）jinzhiqiang@itbb.org.
更新日期/Last Update: 2017-11-03