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来自大白菜(Brassica campestris L. ssp. pekinensis)的两个同源果胶酯酶基因的启动子序列以及该启动子驱动的GUS基因在烟草植株中的花粉特异性表达。

Promoter sequences of two homologous pectin esterase genes from Chinese cabbage (Brassica campestris L. ssp. pekinensis) and pollen-specific expression of the GUS gene driven by a promoter in tobacco plants.

作者信息

Kim H U, Park B S, Jin Y M, Chung T Y

机构信息

Cytogenetics Division, National Institute of Agricultural Science and Technology, Rural Development Administration, Suwon, Korea.

出版信息

Mol Cells. 1997 Feb 28;7(1):21-7.

PMID:9085260
Abstract

The promoter regions of two genomic clones, GBAN215-6 and GBAN215-12 from Chinese cabbage (Brassica campestris L. ssp. pekinensis), were sequenced. The nucleotide sequences of their promoter regions were compared with that of the Bp19 pollen-specific gene of Brassca napus. High nucleotide sequence homologies were observed among these three genes in the region between 210 bp upstream and the putative transcription start site. A sequence motif TGTGGTG, which is similar to that of the PB core motif (TGTGGTT) of two tomato pollen-specific genes, LAT52 and LAT56, was present in these two cloned genes. To determine regulatory sequences responsible for the anther-specific expression of the gene BAN215-6, two recombinant plasmids, pBPE3 (-274- + 109) and pBPE4 (-816- + 109) containing different lengths of the promoter fused with the GUS gene, were constructed and introduced into tobacco plants by Agrobacterium-mediated transformation. The result showed that the 383 bp (-274- + 109) of the BAN215-6 promoter region was sufficient for the anther-specific expression of the GUS gene. The GUS expression in a tobacco plants transformed with these constructs was first detected in uninucleate microspores and persisted at in vitro germinated pollen tubes. The expression level was increased during anther development, reaching the highest level in mature pollens.

摘要

对来自大白菜(Brassica campestris L. ssp. pekinensis)的两个基因组克隆GBAN215 - 6和GBAN215 - 12的启动子区域进行了测序。将它们启动子区域的核苷酸序列与甘蓝型油菜的Bp19花粉特异性基因的序列进行了比较。在这三个基因上游210 bp至假定转录起始位点之间的区域观察到高度的核苷酸序列同源性。在这两个克隆基因中存在一个序列基序TGTGGTG,它与两个番茄花粉特异性基因LAT52和LAT56的PB核心基序(TGTGGTT)相似。为了确定负责基因BAN215 - 6花药特异性表达的调控序列,构建了两个重组质粒pBPE3(-274 - +109)和pBPE4(-816 - +109),它们含有与GUS基因融合的不同长度的启动子,并通过农杆菌介导的转化导入烟草植株。结果表明,BAN215 - 6启动子区域的383 bp(-274 - +109)足以驱动GUS基因的花药特异性表达。用这些构建体转化的烟草植株中,GUS表达首先在单核小孢子中检测到,并在体外萌发的花粉管中持续存在。在花药发育过程中表达水平升高,在成熟花粉中达到最高水平。

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