Colot V, Robert L S, Kavanagh T A, Bevan M W, Thompson R D
Plant Breeding Institute, Maris Lane, Trumpington, Cambridge CB2 2LQ, UK.
EMBO J. 1987;6(12):3559-64. doi: 10.1002/j.1460-2075.1987.tb02685.x.
The developing cereal grain accumulates large quantities of proteins which are unique to the endosperm tissue. The DNA sequences which determine their endosperm-specific expression have not yet been identified. In the absence of a suitable transformation-regeneration system for cereals, we have investigated whether chimaeric genes consisting of low mol. wt (LMW) and high mol. wt (HMW) glutenin gene upstream sequences coupled to the coding region of the bacterial chloramphenicol acetyl transferase (CAT) gene could be specifically expressed in transgenic tobacco. The fusions, made in a Ti-derived binary vector, were introduced into tobacco via Agrobacterium tumefaciens-mediated transformation and their activity assayed. Both the LMW and HMW glutenin chimaeric genes exhibited endosperm-specific CAT activity in the transformed plants. In addition, a deletion series of the LMW glutenin sequence indicated that sequences present between 326 bp and 160 bp upstream of the transcription start point are necessary to confer endosperm-specific CAT activity.
发育中的谷物种子积累了大量胚乳组织特有的蛋白质。决定其胚乳特异性表达的DNA序列尚未被鉴定出来。由于缺乏适合谷物的转化再生系统,我们研究了由低分子量(LMW)和高分子量(HMW)谷蛋白基因上游序列与细菌氯霉素乙酰转移酶(CAT)基因编码区组成的嵌合基因是否能在转基因烟草中特异性表达。在Ti衍生的二元载体中构建的融合基因,通过根癌农杆菌介导的转化导入烟草,并检测其活性。LMW和HMW谷蛋白嵌合基因在转化植株中均表现出胚乳特异性CAT活性。此外,LMW谷蛋白序列的缺失系列表明,转录起始点上游326 bp至160 bp之间的序列是赋予胚乳特异性CAT活性所必需的。