甘蔗UGPase 5′侧翼序列克隆及缺失表达分析

doi:10.7525/j.issn.1673-5102.2014.01.009

植物研究 ›› 2014, Vol. 34 ›› Issue (1): 62-67.doi: 10.7525/j.issn.1673-5102.2014.01.009

甘蔗UGPase 5′侧翼序列克隆及缺失表达分析

叶冰莹^1,2；王婷^1,2；何文锦^1,2；邱思^1,2；陈由强^1,2,3*

1.福建师范大学生命科学学院，福州　350108；2.农业部福建甘蔗生物学与遗传育种重点实验室，福州　350108；3.发育与神经生物学福建省高等学校重点实验室,福州　350108

收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2014-01-20 发布日期:2014-01-20
通讯作者: 陈由强
基金资助:

Cloning and Loss Expression Analysis of 5′ Flanking Sequence of UDP-glucose Pyrophosphorylase from Sugarcane

YE Bing-Ying;WANG Ting;HE Wen-Jin;QIU Si;CHEN You-Qiang;*

1.College of Life Science,Fujian Normal University,Fuzhou　350108；2.Key Lab of Fujian Sugarcane Biology and Genetic Breeding,Ministry of Agriculture,Fuzhou　350108；3.State Key Laboratory of Developmental Biology and Neurobiology,Institution for Higher Learning in Fujian,Fuzhou　350108

Received:1900-01-01 Revised:1900-01-01 Online:2014-01-20 Published:2014-01-20
Contact: CHEN You-
Supported by:

摘要/Abstract

摘要： 以甘蔗(FN95;-1702)为材料，通过接头连接PCR方法克隆该基因不同长度5′侧翼序列。将不同长度的5′侧翼序列连同UGPase基因的外显子片段定向插入到GUS基因上游，在保证其后GUS编码框不发生偏移的情况下，插入的UGPase外显子融合GUS表达成为新的报告基因。根据此策略，构建了一系列表达结构为5′ Flanking Sequence-UGPase Exon-GUS-Nos polyA的5′侧翼序列缺失表达载体，进行启动子活性分析。注射法转染烟草叶片组织检测GUS瞬时表达，分析结果表明，所克隆到的UGPase基因5′端侧翼序列不具有启动子活性。

关键词: 5&prime, 侧翼序列, 缺失表达, 甘蔗, UGPase

Abstract: The different lengths of 5′ flanking sequence of UGPase were cloned by Adaptor-ligation PCR. These different lengths of 5′ flanking sequences of UGPase gene were fused with the coding sequence of GUS (β-glucuronidase) gene to construct fusion genes. All the fusion genes were injected into leaves of Nicotiana tabacum for transient GUS expression. The results showed that the 5′ flanking sequence of UGPase gene did not have any promoter activity.

Key words: 5&prime, flanking sequences;loss expression;Sugarcane;UGPase

中图分类号:

S435.661

叶冰莹;王婷;何文锦;邱思;陈由强;*. 甘蔗UGPase 5′侧翼序列克隆及缺失表达分析[J]. 植物研究, 2014, 34(1): 62-67.

YE Bing-Ying;WANG Ting;HE Wen-Jin;QIU Si;CHEN You-Qiang;*. Cloning and Loss Expression Analysis of 5′ Flanking Sequence of UDP-glucose Pyrophosphorylase from Sugarcane[J]. Bulletin of Botanical Research, 2014, 34(1): 62-67.

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甘蔗UGPase 5′侧翼序列克隆及缺失表达分析

Cloning and Loss Expression Analysis of 5′ Flanking Sequence of UDP-glucose Pyrophosphorylase from Sugarcane

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