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Isolation of the Ankyrin gene promoter and function of its 5UTR intron from tea plant ( Camellia sinensis L.) *(PDF)

Chinese Journal of Applied & Environmental Biology[ISSN:1006-687X/CN:51-1482/Q]

Issue:
2020 01
Page:
1-11
Research Field:
Articles
Publishing date:

Info

Title:
Isolation of the Ankyrin gene promoter and function of its 5UTR intron from tea plant ( Camellia sinensis L.) *
Author(s):
ZHENG Shizhong1 2 JIANG Shengtao1 2 CHEN Meixia1 2 LIN Yuling3 LAI Zhongxiong3** & LIN Jinke 4**
1 College of Life Sciences, Ningde Normal University , Ningde 352100, China
2 Institute of Fujian Higher Education for Local Bio-resources in Ningde City , Ningde 352100, China
3 Institute of Horticultural Biotechnology, Fujian Agriculture and Forestry University , Fuzhou 350002, China 4 Anxi College of Tea Science, Fujian Agriculture and Forestry University , Fuzhou 350002, China
Keywords:
tea plant CsAnkyrin promoter cis-regulatory element 5UTR intron
CLC:
-
PACS:
DOI:
10.19675/j.cnki.1006-687x.2019.05011
DocumentCode:

Abstract:
This study aimed to investigate the regulation in the expression process of CsAnkyrin in tea plant. A 1 010 bp 5’-flanking sequence of CsAnkyrin was isolated from the shoots of tea cultivar ‘1005’ through genome-walking method, designated as proAnk. Sequence analysis was carried out by online bioinformatics resources, such as Neural Network Promotor Prediction and PlantCARE to reveal its basic core promoter, transcription start site and cis-regulatory elements. The results showed, some typical cis-acting elements such as TATA-box, CAAT-box elements, some cis-acting elements responding to hormone, light, stresses, and some other cis-elements with unknown or specific function were contained within proAnk, suggesting that proAnk is an inducible promoter. A 5’UTR intron was also contained in the proAnk sequence. The CsAnkyrin promoter sequences with intron or without intron sequence (+intron/-intron) w ere cloned into the expression vector pCAMBIA1301 to replace CaMV35S promoter and produce a new recombinant expression vector, respectively. The recombinant plasmid was then transformed into Arabidopsis thaliana through floral dip method. The GUS histochemical analysis of transgenic Arabidopsis thaliana revealed that the proAnk including 5’UTR intron can not drive the expression of GUS gene, but the sequences excluding 5’UTR intron can do it. The Semi-Quantitative RT-PCR showed that the 5’UTR intron was not removed during the transcriptional process, but transcripted to mRNA, suggesting that the 5’UTR intron of CsAnkyrin gene negatively regulate the down-stream?gene expression, and may work in the the?process?of translation.

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Last Update: 2019-07-26