Expressed salt stress induced full length-cDNA library of <EM>Salix matsudana</EM> using yeast Invsc1

YANG Ye; JIANG Jing; QIAO Gui-rong; ZHOU Jing; CHEN Yin; HE Zheng-quan; LI Hai-ying; ZHUO Ren-ying

Volume 26 Issue 4

Aug. 2009

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Article Navigation > Journal of Zhejiang A&F University > 2009 > 26(4): 473-478

YANG Ye, JIANG Jing, QIAO Gui-rong, ZHOU Jing, CHEN Yin, HE Zheng-quan, LI Hai-ying, ZHUO Ren-ying. Expressed salt stress induced full length-cDNA library of Salix matsudana using yeast Invsc1[J]. Journal of Zhejiang A&F University, 2009, 26(4): 473-478.

Citation:

YANG Ye, JIANG Jing, QIAO Gui-rong, ZHOU Jing, CHEN Yin, HE Zheng-quan, LI Hai-ying, ZHUO Ren-ying. Expressed salt stress induced full length-cDNA library of Salix matsudana using yeast Invsc1[J]. Journal of Zhejiang A&F University, 2009, 26(4): 473-478.

Expressed salt stress induced full length-cDNA library of Salix matsudana using yeast Invsc1

1.
Biotechnology Research Center / Hubei Province Key Laboratory of Natural Products Research and Development， China Three Gorges University，Yichang 443002，Hubei，China;
2.
Research Institute of Subtropical Forestry，The Chinese Academy of Forestry，Fuyang 311400，Zhejiang，China

More Information

Corresponding author: ZHUO Ren-ying
Publish Date: 2009-08-20

Abstract

The studies of Salix matsudana on salt-resistance molecular mechanism and searching new salt-tolerant relation genes are limited at present. It is necessary to develop drought-tolerant gene using this valuable tree resource，so a salt stress induced full-length cDNA Library of Salix matsudana was constructed using Clontech Laboratories，Inc. SMART cDNA Library Construction Protocol. Total RNA was extracted from S. matsudana under 100 mmol·L-1 NaCl stress for 24 h. After synthesizing the first-strand cDNA，the double-strand cDNA was amplified with the long-distance Polymerase Chain Reaction （LD-PCR）. The result showed that insertion fragment was about 1 000 bp length in the library by PCR. The full-length cDNA fragment was linked to yeast vector pYES2.1 by pYES2.1 TOPO TA Expression Kit；then the linked product was transferred into yeast Invsc1. Finally，clones were selected by SC-U medium containing 80 － 130 g·L－1 NaCl. The experiment shows that the salt-tolerance of transformants yeast improved greatly compared with the wild-type yeast. The results provide an important tool for the study of cloning new salt-tolerance relation genes and the salt tolerance mechanisms of the S. matsudana. ［Ch，6 fig. 17 ref.］
- botany,
- Salix matsudana,
- cDNA library,
- salt tolerance,
- yeast,
- drought tolerance related genes

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

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Expressed salt stress induced full length-cDNA library of Salix matsudana using yeast Invsc1

1. Biotechnology Research Center / Hubei Province Key Laboratory of Natural Products Research and Development， China Three Gorges University，Yichang 443002，Hubei，China;

2. Research Institute of Subtropical Forestry，The Chinese Academy of Forestry，Fuyang 311400，Zhejiang，China

Corresponding author: ZHUO Ren-ying

Publish Date: 2009-08-20

Key words:

Abstract: The studies of Salix matsudana on salt-resistance molecular mechanism and searching new salt-tolerant relation genes are limited at present. It is necessary to develop drought-tolerant gene using this valuable tree resource，so a salt stress induced full-length cDNA Library of Salix matsudana was constructed using Clontech Laboratories，Inc. SMART cDNA Library Construction Protocol. Total RNA was extracted from S. matsudana under 100 mmol·L-1 NaCl stress for 24 h. After synthesizing the first-strand cDNA，the double-strand cDNA was amplified with the long-distance Polymerase Chain Reaction （LD-PCR）. The result showed that insertion fragment was about 1 000 bp length in the library by PCR. The full-length cDNA fragment was linked to yeast vector pYES2.1 by pYES2.1 TOPO TA Expression Kit；then the linked product was transferred into yeast Invsc1. Finally，clones were selected by SC-U medium containing 80 － 130 g·L－1 NaCl. The experiment shows that the salt-tolerance of transformants yeast improved greatly compared with the wild-type yeast. The results provide an important tool for the study of cloning new salt-tolerance relation genes and the salt tolerance mechanisms of the S. matsudana. ［Ch，6 fig. 17 ref.］

Expressed salt stress induced full length-cDNA library of Salix matsudana using yeast Invsc1

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通讯作者: 陈斌, bchen63@163.com

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Expressed salt stress induced full length-cDNA library of Salix matsudana using yeast Invsc1

1. Biotechnology Research Center / Hubei Province Key Laboratory of Natural Products Research and Development， China Three Gorges University，Yichang 443002，Hubei，China;

2. Research Institute of Subtropical Forestry，The Chinese Academy of Forestry，Fuyang 311400，Zhejiang，China

Corresponding author: ZHUO Ren-ying

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Expressed salt stress induced full length-cDNA library of Salix matsudana using yeast Invsc1

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通讯作者: 陈斌, bchen63@163.com

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Expressed salt stress induced full length-cDNA library of Salix matsudana using yeast Invsc1

1. Biotechnology Research Center / Hubei Province Key Laboratory of Natural Products Research and Development， China Three Gorges University，Yichang 443002，Hubei，China; 2. Research Institute of Subtropical Forestry，The Chinese Academy of Forestry，Fuyang 311400，Zhejiang，China

Corresponding author: ZHUO Ren-ying

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1. Biotechnology Research Center / Hubei Province Key Laboratory of Natural Products Research and Development， China Three Gorges University，Yichang 443002，Hubei，China;

2. Research Institute of Subtropical Forestry，The Chinese Academy of Forestry，Fuyang 311400，Zhejiang，China