Tsutsumi R, Tsutsumi K, Numazaki M, Ishikawa K
Eur J Biochem. 1984 Jul 2;142(1):161-4. doi: 10.1111/j.1432-1033.1984.tb08264.x.
Double-stranded DNA was synthesized with reverse transcriptase from size-fractionated poly(A)-containing RNA from rat ascites hepatoma cells. The cDNA was introduced into Escherichia coli HB101 using pBR322 DNA as a cloning vector. Several plasmids containing aldolase A cDNA were identified by colony hybridization with 32P-labeled cDNA prepared from immunologically purified aldolase A mRNA. The partial amino acid sequence of the cDNA sequence was determined, and found to coincide with that of rabbit aldolase A. Using aldolase A cDNA as a hybridization probe, the aldolase A mRNA concentrations in various rat tissues were analysed, and two aldolase A mRNA species differing in nucleotide length were found; the smaller mRNA (about 1550 nucleotides) in muscle, and the larger one (about 1650 nucleotides) in brain and hepatoma cells.
用逆转录酶从大鼠腹水肝癌细胞中经大小分级的含聚腺苷酸（poly(A)）的RNA合成双链DNA。以pBR322 DNA作为克隆载体，将该互补DNA（cDNA）导入大肠杆菌HB101。通过与从经免疫纯化的醛缩酶A信使核糖核酸（mRNA）制备的32P标记的cDNA进行菌落杂交，鉴定出几个含有醛缩酶A cDNA的质粒。测定了该cDNA序列的部分氨基酸序列，发现其与兔醛缩酶A的序列一致。用醛缩酶A cDNA作为杂交探针，分析了各种大鼠组织中醛缩酶A mRNA的浓度，发现了两种核苷酸长度不同的醛缩酶A mRNA；肌肉中较小的mRNA（约1550个核苷酸），以及脑和肝癌细胞中较大的mRNA（约1650个核苷酸）。
