水稻Rubisco激酶基因叶绿体转运肽增强转基因表达

李想; 林智敏; 陈在杰; 王成虎; 刘霞; 王锋

doi:10.19303/j.issn.1008-0384.2013.02.002

水稻Rubisco激酶基因叶绿体转运肽增强转基因表达

doi: 10.19303/j.issn.1008-0384.2013.02.002

李想^1,2,
林智敏²,
陈在杰²,
王成虎²,
刘霞²,
王锋^1,2

1. 福建师范大学生命科学学院,福建福州 350000;
2. 福建省农业科学院生物技术研究所,福建福州,350003

基金项目:

福建省科技计划项目——省属公益类科研院所基本科研专项(2011R1021-3)

详细信息

作者简介:
李想(1982-),女,硕士研究生,主要从事植物分子生物学研究;王锋(1963-),男,博士,研究员,主要从事植物基因工程研究(E-mail:wf@fjage.org)

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出版历程
- 收稿日期: 2012-11-12
- 刊出日期: 2013-02-18

Chloroplast Targeting Signal of a Rice Rubisco Activase Gene Enhances Transgene Expression

1. School of Life Sciences,Fujian Normal University,Fuzhou,Fujian 350000,China;
2. Biotechnology Institute,Fujian Academy of Agricultural Sciences,Fuzhou,Fujian 350003,China

摘要

摘要: 将水稻RCA基因N端预测的转运肽(TP)序列融合报告基因GFP,构建瞬时表达载体pGD-RCATP-GFP,转化农杆菌后,注射侵染烟草叶片,通过荧光显微镜观察瞬时表达情况,结果表明外源蛋白GFP定位在叶绿体上,依此预测水稻RCA基因N端48aa为叶绿体转运肽。将该序列与报告基因GUS融合,构建植物表达载体p1300-RCATP-AGS,转化水稻后,检测阳性株叶片的GUS酶活,其GUS蛋白表达效率是未连有叶绿体转运肽的转基因植物的4.2倍左右。
- RCA(二磷酸核酮糖羧化酶激酶) /
- 叶绿体转运肽 /
- GUS酶活分析
Abstract: N-terminal sequence of the protein encoded by rice RCA gene was predicted to be a chloroplast transit peptide.The 144bp fragment of this gene was fused with GFP to construct a transient expression vector pGD-RCATP-GFP for genetic transformation.Transient transformation of tobacco was produced by the Agrobacterium-mediated infiltration method.Fluorescence microscopy analyses demonstrated that,in cells expressing the RCATP:GFP fusion protein,the fluorescence was specifically targeted to chloroplast.Then this fragment was fused with GUS to construct a vector p1300-RCATP-AGS for genetic transformation.To analyze GUS activity for the leaves of the positive plants,we discovered that the RCA TP sequence increased GUS protein expression levels about 3 fold,when compared to without RCA TP sequence.
- RCA(Rubisco activase) /
- chloroplast transit peptide /
- GUS activity analysis

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参考文献(15)

[1]	梅杨,李海蓝,谢晋,等.核酮糖-1,5-二磷酸羧化酶/加氧酶(Rubisco)[J].植物生理学通讯,2007,43(2):363-368.
[2]	ZHANG G,WANG W,ZOU Q.Molecular biology ofRubisco activase[J].Plant Physio Commu,2004,40(5):633-637.
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[4]	祝钦泷,谢先荣,睦顺照,等.彩叶草Rubisco活化酶基因SsRCAD的分子特征及其表达模式[J].西北植物学报,2011,31(6):1097-1104.
[5]	吴光耀.核编码叶绿体蛋白中的转运肽[J].植物生理学通讯,1991,27(3):166-172.
[6]	YUZO N,AKIRA T,KENICHI KAMIJO.Proteomicanalysis of rice leaf,stem and root tissues during growth course[J].Proteomics,2006,6:3665-3670.
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水稻Rubisco激酶基因叶绿体转运肽增强转基因表达

doi: 10.19303/j.issn.1008-0384.2013.02.002

作者简介:
李想(1982-),女,硕士研究生,主要从事植物分子生物学研究;王锋(1963-),男,博士,研究员,主要从事植物基因工程研究(E-mail:wf@fjage.org)

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Chloroplast Targeting Signal of a Rice Rubisco Activase Gene Enhances Transgene Expression

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水稻Rubisco激酶基因叶绿体转运肽增强转基因表达

doi: 10.19303/j.issn.1008-0384.2013.02.002

作者简介: 李想(1982-),女,硕士研究生,主要从事植物分子生物学研究;王锋(1963-),男,博士,研究员,主要从事植物基因工程研究(E-mail:wf@fjage.org)

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Chloroplast Targeting Signal of a Rice Rubisco Activase Gene Enhances Transgene Expression

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出版历程

目录

作者简介:
李想(1982-),女,硕士研究生,主要从事植物分子生物学研究;王锋(1963-),男,博士,研究员,主要从事植物基因工程研究(E-mail:wf@fjage.org)