Barros M D, Czarnecka E, Gurley W B
Department of Microbiology and Cell Science, University of Florida, Gainesville 32611.
Plant Mol Biol. 1992 Jul;19(4):665-75. doi: 10.1007/BF00026792.
A total of 32 mutations were generated within the TATA-proximal site 1 (-72 to -47) of soybean heat shock gene Gmhsp17.5E in order to functionally define the optimal configuration of sequences within the heat shock element (HSE). Mutants were tested in vivo utilizing sunflower tumors transformed by a T-DNA based vector. Promoter activity was determined by S1 nuclease hybrid protection analysis of tumor transcripts. A total of five repeats (5'-nGAAn-3' or 5'-nTTCn-3') which comprise the HSE at site 1 were required for full transcription induction by heat stress. Analysis of non-conserved bases flanking the central trinucleotide block indicated that 5'-aGAAg'-3' is the optimum sequence for the 5 bp repeat.
为了从功能上确定热激元件（HSE）内序列的最佳构型，在大豆热激基因Gmhsp17.5E的TATA近端位点1（-72至-47）内共产生了32个突变。利用基于T-DNA载体转化的向日葵肿瘤在体内对突变体进行了测试。通过对肿瘤转录本进行S1核酸酶杂交保护分析来确定启动子活性。热应激完全诱导转录需要总共五个重复序列（5'-nGAAn-3'或5'-nTTCn-3'），它们构成了位点1的HSE。对中央三核苷酸块侧翼的非保守碱基分析表明，5'-aGAAg'-3'是5 bp重复序列的最佳序列。
