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Identification and preliminary analysis of a new PCP promoter from Brassica rapa ssp. chinensis

The promoter of Brassica campestris Male Fertile 5 ( BcMF5 ), a pollen coat protein member, class A ( PCP-A ) gene family, was isolated from Brassica rapa L. ssp. chinensis Makino (Chinese cabbage-pak-choi) by Thermal Asymmetric Interlaced Polymerase Chain Reaction (TAIL-PCR). Sequence analysis sugg... Full description

Journal Title: Molecular Biology Reports 2008, Vol.35(4), pp.685-691
Main Author: Zhang, Qiang
Other Authors: Liu, Huizhi , Cao, Jiashu
Format: Electronic Article Electronic Article
Language: English
Subjects:
PCP
ID: ISSN: 0301-4851 ; E-ISSN: 1573-4978 ; DOI: 10.1007/s11033-007-9141-z
Link: http://dx.doi.org/10.1007/s11033-007-9141-z
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recordid: springer_jour10.1007/s11033-007-9141-z
title: Identification and preliminary analysis of a new PCP promoter from Brassica rapa ssp. chinensis
format: Article
creator:
  • Zhang, Qiang
  • Liu, Huizhi
  • Cao, Jiashu
subjects:
  • Brassica rapa ssp.
  • Brassica campestris Male Fertile 5(BcMF5)
  • PCP
  • Promoter
ispartof: Molecular Biology Reports, 2008, Vol.35(4), pp.685-691
description: The promoter of Brassica campestris Male Fertile 5 ( BcMF5 ), a pollen coat protein member, class A ( PCP-A ) gene family, was isolated from Brassica rapa L. ssp. chinensis Makino (Chinese cabbage-pak-choi) by Thermal Asymmetric Interlaced Polymerase Chain Reaction (TAIL-PCR). Sequence analysis suggested that the 605-bp promoter of BcMF5 appears to be a pollen promoter. In an attempt to confirm the promoter activity of BcMF5 promoter, −609 to +3 bp and −377 to +3 bp fragments of the upstream sequence of BcMF5 were inserted at the site upstream of the coding region of the uid A gene in the sense orientation to construct two deletion expression vectors. Transient expression analysis in onion epidermal cells by particle bombardment showed that both −609 to +3 bp and −377 to +3 bp fragments of BcMF5 promoter were capable of driving β-glucuronidase gene expression. Furthermore, by Agrobacterium-mediated genetic transformation method, Arabidopsis transgenic Kan R plants were obtained. GUS assay analysis revealed that the promoter of BcMF5 induced gene expression at the early stage of anther development and drove high levels of GUS expression in anther walls, upper regions of petals, pollen, and pollen tubes in the middle and late stage of anther development, but did not drive any expression in sepals and pistils.
language: eng
source:
identifier: ISSN: 0301-4851 ; E-ISSN: 1573-4978 ; DOI: 10.1007/s11033-007-9141-z
fulltext: fulltext
issn:
  • 1573-4978
  • 15734978
  • 0301-4851
  • 03014851
url: Link


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titleIdentification and preliminary analysis of a new PCP promoter from Brassica rapa ssp. chinensis
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subjectBrassica rapa ssp. ; Brassica campestris Male Fertile 5(BcMF5) ; PCP ; Promoter
descriptionThe promoter of Brassica campestris Male Fertile 5 ( BcMF5 ), a pollen coat protein member, class A ( PCP-A ) gene family, was isolated from Brassica rapa L. ssp. chinensis Makino (Chinese cabbage-pak-choi) by Thermal Asymmetric Interlaced Polymerase Chain Reaction (TAIL-PCR). Sequence analysis suggested that the 605-bp promoter of BcMF5 appears to be a pollen promoter. In an attempt to confirm the promoter activity of BcMF5 promoter, −609 to +3 bp and −377 to +3 bp fragments of the upstream sequence of BcMF5 were inserted at the site upstream of the coding region of the uid A gene in the sense orientation to construct two deletion expression vectors. Transient expression analysis in onion epidermal cells by particle bombardment showed that both −609 to +3 bp and −377 to +3 bp fragments of BcMF5 promoter were capable of driving β-glucuronidase gene expression. Furthermore, by Agrobacterium-mediated genetic transformation method, Arabidopsis transgenic Kan R plants were obtained. GUS assay analysis revealed that the promoter of BcMF5 induced gene expression at the early stage of anther development and drove high levels of GUS expression in anther walls, upper regions of petals, pollen, and pollen tubes in the middle and late stage of anther development, but did not drive any expression in sepals and pistils.
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titleIdentification and preliminary analysis of a new PCP promoter from Brassica rapa ssp. chinensis
descriptionThe promoter of Brassica campestris Male Fertile 5 ( BcMF5 ), a pollen coat protein member, class A ( PCP-A ) gene family, was isolated from Brassica rapa L. ssp. chinensis Makino (Chinese cabbage-pak-choi) by Thermal Asymmetric Interlaced Polymerase Chain Reaction (TAIL-PCR). Sequence analysis suggested that the 605-bp promoter of BcMF5 appears to be a pollen promoter. In an attempt to confirm the promoter activity of BcMF5 promoter, −609 to +3 bp and −377 to +3 bp fragments of the upstream sequence of BcMF5 were inserted at the site upstream of the coding region of the uid A gene in the sense orientation to construct two deletion expression vectors. Transient expression analysis in onion epidermal cells by particle bombardment showed that both −609 to +3 bp and −377 to +3 bp fragments of BcMF5 promoter were capable of driving β-glucuronidase gene expression. Furthermore, by Agrobacterium-mediated genetic transformation method, Arabidopsis transgenic Kan R plants were obtained. GUS assay analysis revealed that the promoter of BcMF5 induced gene expression at the early stage of anther development and drove high levels of GUS expression in anther walls, upper regions of petals, pollen, and pollen tubes in the middle and late stage of anther development, but did not drive any expression in sepals and pistils.
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abstractThe promoter of Brassica campestris Male Fertile 5 ( BcMF5 ), a pollen coat protein member, class A ( PCP-A ) gene family, was isolated from Brassica rapa L. ssp. chinensis Makino (Chinese cabbage-pak-choi) by Thermal Asymmetric Interlaced Polymerase Chain Reaction (TAIL-PCR). Sequence analysis suggested that the 605-bp promoter of BcMF5 appears to be a pollen promoter. In an attempt to confirm the promoter activity of BcMF5 promoter, −609 to +3 bp and −377 to +3 bp fragments of the upstream sequence of BcMF5 were inserted at the site upstream of the coding region of the uid A gene in the sense orientation to construct two deletion expression vectors. Transient expression analysis in onion epidermal cells by particle bombardment showed that both −609 to +3 bp and −377 to +3 bp fragments of BcMF5 promoter were capable of driving β-glucuronidase gene expression. Furthermore, by Agrobacterium-mediated genetic transformation method, Arabidopsis transgenic Kan R plants were obtained. GUS assay analysis revealed that the promoter of BcMF5 induced gene expression at the early stage of anther development and drove high levels of GUS expression in anther walls, upper regions of petals, pollen, and pollen tubes in the middle and late stage of anther development, but did not drive any expression in sepals and pistils.
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doi10.1007/s11033-007-9141-z
pages685-691
date2008-12