CLONING AND SEQUENCE-ANALYSIS OF A CDNA-ENCODING PORCINE MITOCHONDRIAL ASPARTATE-AMINOTRANSFERASE PRECURSOR
CLONING AND SEQUENCE-ANALYSIS OF A CDNA-ENCODING PORCINE MITOCHONDRIAL ASPARTATE-AMINOTRANSFERASE PRECURSOR
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DOI：
10.1073/pnas.82.18.6065
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发表时间：
1985-01-01
影响因子：
11.1
通讯作者：
MORINO, Y
中科院分区：
文献类型：
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作者：
JOH, T;NOMIYAMA, H;MORINO, Y
根据猪线粒体天冬氨酸氨基转移酶(mAspATase；L-天冬氨酸：2-氧戊二酸氨基转移酶，EC2.6.1.1)前体基因的序列推导出其一级结构。应用Okayama和Berg[Okayama，H.和Berg，P.(1982)Mol]的载体/引物法，构建了猪肝脏Poly(A)+RNA的cDNA克隆文库。牢房。比奥尔。2、161-170]。使用8个寡脱氧核苷酸的混合物作为探针对文库进行猪mAspATase序列的筛选。该探针的序列来源于猪mAspATase残基196-201的已知氨基酸序列。从序列分析中筛选出两个含有约2500和2600个碱基对的插入片段的重组质粒。从该序列预测的氨基酸序列表明，猪mAspATase前体由401个氨基酸残基组成的成熟酶和29个氨基酸残基的氨基末端片段组成，该前序列含有4个碱性氨基酸残基，没有酸性残基，也没有疏水性氨基酸延伸。将该29个氨基酸的mAspATase前体片段的序列与其他线粒体酶的前序列进行了比较。
The primary structure of pig mitochondrial aspartate aminotransferase (mAspATase; L-aspartate:2-oxoglutarate aminotransferase, EC 2.6.1.1) precursor was deduced from its cDNA sequence. A library of cDNA clones was constructed from pig liver poly(A)+ RNA by applying the vector/primer method of Okayama and Berg [Okayama, H. and Berg, P. (1982) Mol. Cell. Biol. 2, 161-170]. The library was screened for pig mAspATase sequences by using a mixture of eight oligodeoxyribonucleotides as a probe. The sequences of the probe were deduced from the known amino acid sequence of pig mAspATase residues 196-201. Two recombinant plasmids containing inserts of about 2500 and 2600 base pairs were selected from sequence analysis. The amino acid sequence predicted from the cDNA sequence shows that the pig mAspATase precursor consists of the mature enzyme of 401 amino acid residues and an amino-terminal segment of 29 amino acid residues called the "presequence" that contains four basic amino acid residues, no acidic residues, and no hydrophobic amino acid stretch. The sequence of this 29-amino acid mAspATase precursor segment was compared with the presequences of other mitochondrial enzymes.