Koiwa Hisashi, Fukudome Akihito
a Vegetable and Fruit Improvement Center, Department Horticultural Sciences , Texas A&M University , College Station , TX , USA.
b Molecular and Environmental Plant Science Program, Texas A&M University , College Station , TX , USA.
Plant Signal Behav. 2017 Aug 3;12(8):e1346767. doi: 10.1080/15592324.2017.1346767. Epub 2017 Jul 10.
Forward genetic screening of mutants using firefly luciferase (LUC) reporter gene became a standard practice in plant research. Such screenings frequently identified alleles of CPL1 (Carboxyl-terminal Phosphatase-Like 1) regardless of promoters or pathways studied. Expression of the corresponding endogenous genes often shows the minimal difference between wild type and cpl1. Here we show that the LUC coding sequence is responsible for the high expression in cpl1, using a classical RD29a-LUC. Deletion of the LUC 3'-UTR did not change hyperactivation of LUC in cpl1. However, a codon-modified LUC (LUC2) produced similar expression levels both in wild type and in cpl1. These results indicate that the coding region of LUC is responsible for the cpl1-specific LUC overexpression uncoupled with the expression of the endogenous counterpart.
利用萤火虫荧光素酶（LUC）报告基因对突变体进行正向遗传筛选已成为植物研究中的标准做法。无论研究的启动子或途径如何，此类筛选经常鉴定出CPL1（羧基末端磷酸酶样1）的等位基因。相应内源基因的表达在野生型和cpl1之间通常显示出最小差异。在这里，我们使用经典的RD29a-LUC表明，LUC编码序列是cpl1中高表达的原因。LUC 3'-UTR的缺失并未改变cpl1中LUC的超激活。然而，密码子修饰的LUC（LUC2）在野生型和cpl1中均产生相似的表达水平。这些结果表明，LUC的编码区是cpl1特异性LUC过表达的原因，且与内源对应物的表达无关。
