Sakimura K, Kushiya E, Obinata M, Odani S, Takahashi Y
Proc Natl Acad Sci U S A. 1985 Nov;82(21):7453-7. doi: 10.1073/pnas.82.21.7453.
The cDNAs to mRNA for rat gamma gamma enolase (neuron-specific enolase; NSE; EC 4.2.1.11) were isolated from a cDNA library by using differential colony hybridization and a hybrid-selected translation assay. By overlapping of the nucleotide sequences of several cDNA inserts, it was found that they spanned 2232 base pairs (bp) which included 1299 bp of the complete coding region, 68 bp of the 5' noncoding region, and 848 bp of the 3' noncoding region, including a polyadenylylation signal. In addition, the poly(A) tail was also found. The amino acid sequence deduced from the nucleotide sequence was composed of 433 amino acids. Southern blot analysis with a cDNA insert detected one hybridizing fragment in rat genomic DNA digested with several different restriction enzymes. Dot-blot and transfer hybridization analyses of poly(A)+ RNA from developing rat brains showed an increase of NSE mRNA 10-30 days after birth.
通过差异菌落杂交和杂交选择翻译分析,从大鼠γγ烯醇化酶(神经元特异性烯醇化酶;NSE;EC 4.2.1.11)的mRNA的cDNA文库中分离出cDNA。通过重叠几个cDNA插入片段的核苷酸序列,发现它们跨度为2232个碱基对(bp),其中包括1299 bp的完整编码区、68 bp的5'非编码区和848 bp的3'非编码区,包括一个多聚腺苷酸化信号。此外,还发现了多聚(A)尾。从核苷酸序列推导的氨基酸序列由433个氨基酸组成。用cDNA插入片段进行的Southern印迹分析在几种不同限制酶消化的大鼠基因组DNA中检测到一个杂交片段。对发育中的大鼠大脑的聚(A)+RNA进行斑点印迹和转移杂交分析表明,出生后10 - 30天NSE mRNA增加。