Structural analysis of aldolase C gene and the identification of its expressed cell in brain

分离大鼠醛缩酶C基因。该基因由9个被8个内含子打断的外显子组成。测定了该基因的核苷酸序列，包括5‘侧链的801个核苷酸（nt.），基因区3590个核苷酸（nt.）和3’侧链的461个核苷酸（nt.）。通过S1定位和引物延伸的方法确定了两个转录起始位点。醛缩酶cmrna在脑组织和肝癌组织中均有表达。为了鉴定在脑组织中表达醛缩酶C mRNA的细胞，我们用核糖探针对脑和脊髓进行了原位杂交。我们搜索了大脑皮层、小脑、海马体、丘脑。令人惊讶的是，银颗粒在小脑浦肯野细胞和背根神经节和脑核中积累。脊髓外侧运动肌。我们还测量了基因组中该基因的拷贝数。从基因的测量，通过南方印迹滴定，我们得出结论，一个拷贝的大鼠醛缩酶C基因发生在每个大鼠单倍体基因组。

Rat aldolase C gene was isolated. The gene comprises 9 exons interruped by 8 introns. The nucleotide sequence of the gene was determined, which includes 801 nucleotides(nt.) of 5'flanking, 3590 nt. of the gene region and 461 nt. of 3' flanking sequences. Two transcriptional initiation sites were also determined by the method of S1 mapping and primer extension. Aldolase C mRNA was exclusively expressed in brain and some in hepatoma. To identify the cell expressing aldolase C mRNA in brain we performed in situ hypridization of brain and spinal cord using riboprobe. We searched cerebral cortex, cerebellum, hippocampus, thalamus of the brain. Surprisingly silver grains accumulated in the Purkinje cell of the cerebellum and some in dorsal root ganglion and Nuc. motorius lateralis of the spinal cord. We also measured the copy number of the gene in the genome. From the measurement of the geme, titrated by Southern blotting, we conclude that a single copy of the rat aldolase C gene occurs per rat haploid genome.