Lobach D F, Rochelle J M, Watson M L, Seldin M F, Blackshear P J
Howard Hughes Medical Institute Laboratories, Section of Diabetes and Metabolism, Division of Endocrinology, Metabolism, and Nutrition, Department of Medicine.
Genomics. 1993 Jul;17(1):194-204. doi: 10.1006/geno.1993.1301.
We isolated and characterized a genomic clone for the mouse MARCKS-related protein, or MRP, also known as F52 and MacMARCKS. A 3699-bp plasmid contained 407 bp of 5'-flanking region, 186 bp of 5'-untranslated region, 600 bp of protein coding region, 784 bp of a single intron, 746 bp of 3'-untranslated region, and 976 bp of 3'-flanking region. The position of the single intron was identical to the intron position in all known MARCKS mRNAs. When the plasmid containing the genomic sequences was transfected into fibroblasts lacking endogenous Mrp expression, the 407 bp of promoter conferred high-level expression of the full-length, spliced mRNA. The putative promoter was therefore functional; however, despite tissue-specific regulation and transcriptional induction in some cells in a manner similar to that seen with MARCKS expression, the promoters were highly dissimilar at the level of primary sequence (37% identity over 407 bp). Mrp mapped to a position on mouse chromosome 4 that was closely linked to the Lck locus. Numerous additional species that hybridized to the MRP cDNA were noted on Southern blotting of mouse genomic DNA. Five related loci were labeled Mrp-rs1 through Mrp-rs5 (for Mrp-related sequences) and were mapped to mouse chromosomes 10, 17, 15, 13, and X, respectively. Three of these related sequences have been cloned, and all appear to represent pseudogenes.
我们分离并鉴定了小鼠MARCKS相关蛋白（MRP，也称为F52和MacMARCKS）的基因组克隆。一个3699 bp的质粒包含407 bp的5'侧翼区、186 bp的5'非翻译区、600 bp的蛋白质编码区、784 bp的单个内含子、746 bp的3'非翻译区和976 bp的3'侧翼区。单个内含子的位置与所有已知MARCKS mRNA中的内含子位置相同。当将包含基因组序列的质粒转染到缺乏内源性Mrp表达的成纤维细胞中时，407 bp的启动子赋予全长剪接mRNA高水平表达。因此，推定的启动子具有功能；然而，尽管在某些细胞中存在组织特异性调控和转录诱导，其方式与MARCKS表达所见相似，但启动子在一级序列水平上高度不同（407 bp上的同一性为37%）。Mrp定位于小鼠4号染色体上与Lck基因座紧密连锁的位置。在小鼠基因组DNA的Southern印迹中发现了许多与MRP cDNA杂交的其他物种。五个相关基因座分别标记为Mrp-rs1至Mrp-rs5（代表Mrp相关序列），并分别定位于小鼠10、17、15、13和X染色体上。其中三个相关序列已被克隆，似乎都代表假基因。
