Guan L, Scandalios J G
Department of Genetics, North Carolina State University, Raleigh 27695-7614.
Plant J. 1993 Apr;3(4):527-36. doi: 10.1046/j.1365-313x.1993.03040527.x.
A genomic clone containing the maize Cat1 gene has been isolated and its complete DNA sequence determined. The start of transcription has been mapped by primer extension. Six introns were identified in the Cat1 coding region. In order to determine the tissue-specific expression pattern of the Cat1 gene, promoter-reporter gene fusion constructs were made consisting of 2.5 kb and 0.8 kb of the 5' Cat1 sequence fused to the coding region of the beta-glucuronidase (GUS) gene. These fusion constructs were introduced into Nicotiana tabacum cv. Burley 21 and the expression of Cat1-GUS in various tissues was examined. In transgenic tobacco, the Cat1 promoter can drive GUS expression at relatively high levels in mature seeds. GUS activity starts to accumulate at about 10 days after flowering, reaching a maximum at about 22 days after flowering, and decreases thereafter, but persists until after seed desiccation through early germination. Low levels of GUS activity can be detected in pollen. This corresponds to the Cat1 expression pattern observed in maize.
一个包含玉米Cat1基因的基因组克隆已被分离出来,并测定了其完整的DNA序列。转录起始位点已通过引物延伸法进行了定位。在Cat1编码区鉴定出六个内含子。为了确定Cat1基因的组织特异性表达模式,构建了启动子-报告基因融合体,由2.5 kb和0.8 kb的5'端Cat1序列与β-葡萄糖醛酸酶(GUS)基因的编码区融合而成。这些融合构建体被导入烟草品种Burley 21中,并检测了Cat1-GUS在各种组织中的表达。在转基因烟草中,Cat1启动子能够驱动GUS在成熟种子中相对高水平的表达。GUS活性在开花后约10天开始积累,在开花后约22天达到最大值,此后下降,但一直持续到种子干燥至早期萌发。在花粉中可检测到低水平的GUS活性。这与在玉米中观察到的Cat1表达模式一致。