利用酿酒酵母表达氯化钠胁迫下旱柳全长cDNA文库

杨晔; 蒋晶; 乔桂荣; 周婧; 陈银; 何正权; 李海营; 卓仁英

全国中文核心期刊
中国科技核心期刊
CSCD核心库来源期刊
中国农林核心期刊
RCCSE中国核心学术期刊

ISSN 2095-0756 CN 33-1370/S

留言板

尊敬的读者、作者、审稿人, 关于本刊的投稿、审稿、编辑和出版的任何问题, 您可以本页添加留言。我们将尽快给您答复。谢谢您的支持!

姓名
邮箱
手机号码
标题
留言内容
验证码

利用酿酒酵母表达氯化钠胁迫下旱柳全长cDNA文库

杨晔 , 蒋晶 , 乔桂荣 , 周婧 , 陈银 , 何正权 , 李海营 , 卓仁英

文章导航 > 浙江农林大学学报 > 2009 > 26(4): 473-478

杨晔, 蒋晶, 乔桂荣, 等. 利用酿酒酵母表达氯化钠胁迫下旱柳全长cDNA文库[J]. 浙江农林大学学报, 2009, 26(4): 473-478.

引用本文:

杨晔, 蒋晶, 乔桂荣, 等. 利用酿酒酵母表达氯化钠胁迫下旱柳全长cDNA文库[J]. 浙江农林大学学报, 2009, 26(4): 473-478.

YANG Ye, JIANG Jing, QIAO Gui-rong, et al. 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, et al. 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.

利用酿酒酵母表达氯化钠胁迫下旱柳全长cDNA文库

杨晔^1,2,
蒋晶²,
乔桂荣²,
周婧²,
陈银²,
何正权¹,
李海营²,
卓仁英^2,

1.
三峡大学生物技术研究中心/天然产物研究与利用湖北省重点实验室，湖北宜昌 443002;
2.
中国林业科学研究院亚热带林业研究所，浙江富阳 311400

详细信息

通信作者: 卓仁英

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

摘要: 为了研究旱柳Salix matsudana高耐盐无性系I?鄄32的耐盐机制，分离其耐盐相关基因，实验以筛选出的高耐盐旱柳无性系I-32为材料，盐胁迫后提取总核糖核酸（RNA），纯化mRNA，利用改良SMART（switching mechanism at 5’end of RNA transcript）技术合成了全长cDNA，回收500 bp以上cDNA大片段克隆到大肠杆菌Escherichia coli和酵母穿梭载体pYES2.1中，建成旱柳全长cDNA文库，对随机挑取的阳性克隆进行聚合酶链式反应（PCR）鉴定，插入片断平均值为1 000 bp左右，说明所构建的文库达到了用于目的基因分离筛选和表达的建库要求。通过尿嘧啶缺陷型培养基添加80~130 g·L－1氯化钠筛选文库，与野生型酵母菌株相比，转化子的耐盐能力提高。研究结果为克隆与旱柳抗旱相关基因奠定了基础。图6参17
- 植物学 /
- 旱柳 /
- cDNA文库 /
- 耐盐性 /
- 酵母 /
- 抗旱相关基因
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
链接本文:
https://zlxb.zafu.edu.cn/article/id/463

https://zlxb.zafu.edu.cn/article/zjnldxxb/2009/4/473

点击查看大图

计量

文章访问数: 4125
HTML全文浏览量: 198
PDF下载量: 297
被引次数: 0

利用酿酒酵母表达氯化钠胁迫下旱柳全长cDNA文库

1. 三峡大学生物技术研究中心/天然产物研究与利用湖北省重点实验室，湖北宜昌 443002;

2. 中国林业科学研究院亚热带林业研究所，浙江富阳 311400

通信作者: 卓仁英

刊出日期: 2009-08-20

关键词:

摘要: 为了研究旱柳Salix matsudana高耐盐无性系I?鄄32的耐盐机制，分离其耐盐相关基因，实验以筛选出的高耐盐旱柳无性系I-32为材料，盐胁迫后提取总核糖核酸（RNA），纯化mRNA，利用改良SMART（switching mechanism at 5’end of RNA transcript）技术合成了全长cDNA，回收500 bp以上cDNA大片段克隆到大肠杆菌Escherichia coli和酵母穿梭载体pYES2.1中，建成旱柳全长cDNA文库，对随机挑取的阳性克隆进行聚合酶链式反应（PCR）鉴定，插入片断平均值为1 000 bp左右，说明所构建的文库达到了用于目的基因分离筛选和表达的建库要求。通过尿嘧啶缺陷型培养基添加80~130 g·L－1氯化钠筛选文库，与野生型酵母菌株相比，转化子的耐盐能力提高。研究结果为克隆与旱柳抗旱相关基因奠定了基础。图6参17

English Abstract

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

Keywords:

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.］

杨晔, 蒋晶, 乔桂荣, 等. 利用酿酒酵母表达氯化钠胁迫下旱柳全长cDNA文库[J]. 浙江农林大学学报, 2009, 26(4): 473-478.

引用本文:

杨晔, 蒋晶, 乔桂荣, 等. 利用酿酒酵母表达氯化钠胁迫下旱柳全长cDNA文库[J]. 浙江农林大学学报, 2009, 26(4): 473-478.

YANG Ye, JIANG Jing, QIAO Gui-rong, et al. 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:

全文HTML

下载: 全尺寸图片幻灯片

返回文章

用微信扫码二维码

分享至好友和朋友圈

留言板

利用酿酒酵母表达氯化钠胁迫下旱柳全长cDNA文库

通信作者: 卓仁英

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

Corresponding author: ZHUO Ren-ying

计量

出版历程

利用酿酒酵母表达氯化钠胁迫下旱柳全长cDNA文库

1. 三峡大学 生物技术研究中心/天然产物研究与利用湖北省重点实验室，湖北 宜昌 443002; 2. 中国林业科学 研究院 亚热带林业研究所，浙江 富阳 311400

通信作者: 卓仁英

English Abstract

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

全文HTML

目录

1. 三峡大学生物技术研究中心/天然产物研究与利用湖北省重点实验室，湖北宜昌 443002;

2. 中国林业科学研究院亚热带林业研究所，浙江富阳 311400

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