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[1]周冀明,卫志明,刘世贵,等.GUS基因在诸葛菜子叶原生质体中的瞬间表达[J].应用与环境生物学报,1995,1(01):7-11.
 Wei Zhiming,Zhou Jiming,Liu Shigui,et al.TRANSIENT EXPRESSION OF GUS GENE IN COTYLEDEN PROTOPLASTS OF ORYCHOPHRAGMUS VIOLACEUS[J].Chinese Journal of Applied & Environmental Biology,1995,1(01):7-11.
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GUS基因在诸葛菜子叶原生质体中的瞬间表达()
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《应用与环境生物学报》[ISSN:1006-687X/CN:51-1482/Q]

卷:
1卷
期数:
1995年01期
页码:
7-11
栏目:
研究论文
出版日期:
1995-02-25

文章信息/Info

Title:
TRANSIENT EXPRESSION OF GUS GENE IN COTYLEDEN PROTOPLASTS OF ORYCHOPHRAGMUS VIOLACEUS
作者:
周冀明2 卫志明1 刘世贵2 罗鹏3
1. 中国科学院上海植物生理研究所植物分子遗传国家重点实验室, 上海 200032;
2. 四川大学生物工程系, 四川成都 6100641;
3. 四川大学生物系, 四川成都 6100641
Author(s):
Wei Zhiming2 Zhou Jiming1 Liu Shigui2 Luo Peng3
1. National Laboratory of Plant Molecular Genetics of Shanghai Institute of Plant Physiology, Academia Sinica, Shanghai 200032;
2. Depentment of Biotechnology of Sichuan University, Chengdu 610064;
3. Depentment of Biology of Sichuan University, Chengdu 610064
关键词:
诸葛菜原生质体GUS基因瞬间表达
Keywords:
Orychophragmus violaceusprotoplastGUS transient expression
摘要:
本文以诸葛菜无菌苗子叶组织为材料,采用原生质体培养获得成功,建立了原生质体培养高频再生体系,植板率3%,植株再生频率100%.在此基础上,本文首次研究了PEG介导转化诸葛菜原生质体的影响因素,系统地试验了PEG法转化子叶原生质体的过程,发现:最适质粒量为25-30μg;PEG终浓度为15%,pH8.0;另外,表达效率还与质粒的大小有关,较小的质粒具有相对较高的转化效率;而CT-DNA以及热击处理未见有明显的促进作用.
Abstract:
This paper presents the study on the transient expression level of the GUS reporter gene with cotyleden protoplasts of Orychophragmus violaceus. The highest expression level was obtained when a smaller (5.7 kb)plasmid 25-30 μg, 15% and pH 8.0 of PEG were used. The carrier DNA, ctDNA and heat shock treatment seemed no effect on its expression.

参考文献/References:

[1] 王善平等.利用PEG法对GUS基因在杨树原生质体中瞬间表达的研究.实验生物学报.1991.24(1):71-74
[2] 徐晓昕等.诸葛菜叶片原生质体培养.植物生理学报·1998,l4(2):170-174
[3] 许智宏等.植物生理与分子生物学.北京:科学出版社,1987
[4] 周太炎等.中国植物志(第33卷).北京:科学出版社,1987
[5] 罗科等.诸葛菜叶柄原生质体培养再生植株.生物工程学报·1992.8(2):174-177
[6] Ballas N et a1. Transient expression of the plasmid pCaMVCAT in plant protoplasts following transformation with polyerhylene glycol. Rrp Cell Res. 1987,170; 228-234
[7] Prols M et al. Transient gene expression in tobacco protoplasts. Plant Cell Res. 1988,7;221-224
[8] Jefferson AR. Assaying chimeric gene in plants;the GUS gene fusion system. Plnul MolecaLrr Rwlogy Reporter. 1987,4:387-405
[9] Huang Jianqiu el n1. Transient gene expression in protoplasts of soybean. Craekc Mampulakon in Plants. 1991,7(1);53-59
[10] Kohlet F et al. Stable transformation of moth bean Vigan acomtifolin via direct gene transfer. Plant Cell Rep.,1987,6:313-317
[11] Krens AF et al.In vitro transformation of plant protoplasts with Ti-plasmid DNA.Nature.1982.296;72-74

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备注/Memo

备注/Memo:
收稿日期:1994-8-16;接受日期:1994-11-21。
更新日期/Last Update: 1900-01-01