Porsch P, Merkelbach S, Gehlen J, Fladung M
Max-Planck-Institute for Plant Breeding, Department of Plant Breeding and Yield Physiology, Köln, Germany.
Anal Biochem. 1993 May 15;211(1):113-6. doi: 10.1006/abio.1993.1240.
Four different promoters--the 35S, a double 35S promoter, and two different promoter fragments of the ribulose-bisphosphate carboxylase small subunit (rbcS)--were fused to the reporter gene chloramphenicol acetyltransferase (CAT) and transient expression studies were performed in potato protoplasts. Promoter strength was evaluated by using a nonradioactive CAT immunoassay not previously tested with plant cells. The activities of the rbcS promoters were 10-fold less than those of the 35S promoters. Unspecific reactions due to the immunoassay adopted were very low, and the sensitivity was in the range of that of other radioactive and nonradioactive reporter gene assays. The CAT-enzyme-linked immunosorbent assay test is a suitable nonradioactive alternative to test relatively weak promoters in plant systems.
将四个不同的启动子——35S启动子、双35S启动子以及核酮糖-1,5-二磷酸羧化酶小亚基（rbcS）的两个不同启动子片段——与报告基因氯霉素乙酰转移酶（CAT）融合，并在马铃薯原生质体中进行瞬时表达研究。使用一种此前未在植物细胞中测试过的非放射性CAT免疫测定法评估启动子强度。rbcS启动子的活性比35S启动子低10倍。所采用的免疫测定法引起的非特异性反应非常低，其灵敏度与其他放射性和非放射性报告基因测定法相当。CAT酶联免疫吸附测定试验是在植物系统中检测相对较弱启动子的一种合适的非放射性替代方法。
