EXPRESSION OF A TUBER-SPECIFIC STORAGE PROTEIN IN TRANSGENIC TOBACCO PLANTS - DEMONSTRATION OF AN ESTERASE-ACTIVITY
EXPRESSION OF A TUBER-SPECIFIC STORAGE PROTEIN IN TRANSGENIC TOBACCO PLANTS - DEMONSTRATION OF AN ESTERASE-ACTIVITY
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DOI：
10.1002/j.1460-2075.1987.tb02348.x
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发表时间：
1987-05-01
期刊：
影响因子：
11.4
通讯作者：
WILLMITZER, L
中科院分区：
文献类型：
--
作者：
ROSAHL, S;SCHELL, J;WILLMITZER, L
由马铃薯叶/茎特异性表达基因 ST-LS1 的 5'' 上游区域和马铃薯块茎的主要储存蛋白 patatin 的 RNA 编码和 3'' 下游区域组成的嵌合基因，已使用农杆菌系统转移到烟草植物中。该基因的引入导致了 42 kd 大蛋白的叶/茎特异性表达，该蛋白与 patatin 抗血清发生免疫交叉反应。在转基因烟叶中只能检测到少量的较小尺寸的免疫反应蛋白，这表明patatin蛋白在这种异源环境中相当稳定。使用 patatin 特异性探针在转基因烟草叶中检测到的蛋白质大小以及 RNA 大小表明 patatin RNA 在烟草的叶和茎组织中均得到了准确的加工。 patatin 基因的表达导致转化烟草中出现新的酯酶活性，该活性与与 patatin 抗血清发生免疫反应的蛋白质共同迁移。因此，这些数据表明马铃薯蛋白除了充当储存蛋白之外还表现出酶活性。
A chimaeric gene composed of the 5'' upstream region of ST-LS1, a leaf/stem specifically expressed gene from Solanum tuberosum, and the RNA-coding as well as the 3'' downstream region of patatin, the major storage protein of potato tubers, has been transferred into tobacco plants using the Agrobacterium system. The introduction of the gene led to a leaf/stem specific expression of a 42-kd large protein which immunocrossreacts with patatin antiserum. Only low amounts of immunoreacting protein of smaller size could be detected in transgenic tobacco leaves indicating that the patatin protein is fairly stable in this heterologous environment. The size of the protein as well as the size of the RNA detected in transgenic tobacco leaves using a patatin-specifc probe indicates that the patatin RNA was accurately processed in both leaf and stem tissue of tobacco. The expression of the patatin gene led to the appearance of a new esterase activity in the transformed tobacco which co-migrated with a protein immunoreacting with patatin antiserum. These data therefore demonstrate that patatin in addition to serving as a storage protein displays an enzymatic activity.