Cloning and sequence analysis of cDNA for human argininosuccinate lyase.
Cloning and sequence analysis of cDNA for human argininosuccinate lyase.
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人精氨酸琥珀酸裂合酶 cDNA 的克隆和序列分析。
DOI：
10.1073/pnas.83.19.7211
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发表时间：
1986
影响因子：
11.1
通讯作者：
Adcock,M
中科院分区：
文献类型：
--
作者：
O'Brien,WE;McInnes,R;Kalumuck,K;Adcock,M
使用对氨基琥珀酸裂解酶（EC 4.3.2.1）特异的抗体，我们通过筛选在λ gt 11表达载体中构建的人肝cDNA文库来分离两个cDNA克隆。通过质粒选择mRNA的体外翻译证实了这些分离株的身份。其中一个分离株用于重新筛选cDNA文库，并鉴定了一个1565碱基对（bp）的克隆。测定了该克隆的整个核苷酸序列。鉴定了一个开放阅读框，其编码463个氨基酸的蛋白质，预测分子量为51，663。该克隆含有115 bp的5'端非翻译序列和46 bp的3'端非翻译序列。在3'端，距poly（A）序列起始处16 bp处存在一个典型的poly（A）添加位点。比较推导的氨基酸序列的人酶与酵母酶的同源性为56%，当保守的氨基酸变化被考虑在内。酵母蛋白也是463个氨基酸长，分子量为51，944。通过使用来自人-中国仓鼠体细胞杂种的基因组DNA板，将人基因定位于7号染色体。在22号染色体上发现了另一个杂交区，对应于cDNA的5'端的一部分。
Using antibodies specific for argininosuccinate lyase (EC 4.3.2.1), we isolated two cDNA clones by screening a human liver cDNA library constructed in the lambda gt11 expression vector. The identity of these isolates was confirmed by in vitro translation of plasmid-selected mRNA. One of these isolates was used to rescreen the cDNA library and a 1565-base-pair (bp) clone was identified. The entire nucleotide sequence of this clone was determined. An open reading frame was identified which encoded a protein of 463 amino acids with a predicted molecular weight of 51,663. The clone included 115 bp of 5' untranslated sequence and 46 bp of 3' untranslated sequence. A canonical poly(A) addition site was present in the 3' end, 16 bp from the beginning of the poly(A) tract. Comparison of the deduced amino acid sequence of the human enzyme with that of the yeast enzyme revealed a 56% homology, when conservative amino acid changes were taken into consideration. The yeast protein is also 463 amino acids long, with a molecular weight of 51,944. By use of a genomic DNA panel from human-Chinese hamster somatic cell hybrids, the human gene was mapped to chromosome 7. Another hybridizing region, corresponding to a portion of the 5' end of the cDNA, was found on chromosome 22.