Isolation, nucleotide sequence, and expression of a cDNA encoding pig citrate synthase.
Isolation, nucleotide sequence, and expression of a cDNA encoding pig citrate synthase.
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编码猪柠檬酸合酶的 cDNA 的分离、核苷酸序列和表达。
DOI：
10.1021/bi00413a015
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发表时间：
1988
期刊：
影响因子：
2.9
通讯作者：
P. Srere
中科院分区：
文献类型：
--
作者：
Claudia T. Evans;Daniel D. Owens;Balazs Sumegi;G. Kispál;P. Srere
Citrate synthase is a key enzyme of the Krebs tricarboxylic acid cycle and catalyzes the stereospecific synthesis of citrate from acetyl coenzyme A and oxalacetate. The amino acid sequence and three-dimensional structure of pig citrate synthase dimers are known, and regions of the enzyme involved in substrate binding and catalysis have been identified. A cloned complementary DNA sequence encoding pig citrate synthase has been isolated from a pig kidney lambda gt11 cDNA library after screening with a synthetic oligonucleotide probe. The complete nucleotide sequence of the 1.5-kilobase cDNA was determined. The coding region consists of 1395 base pairs and confirms the amino acid sequence of purified pig citrate synthase. The derived amino acid sequence of pig citrate synthase predicts the presence of a 27 amino acid N-terminal leader peptide whose sequence is consistent with the sequences of other mitochondrial signal peptides. A conserved amino acid sequence in the mitochondrial leader peptides of pig citrate synthase and yeast mitochondrial citrate synthase was identified. To express the pig citrate synthase cDNA in Escherichia coli, we employed the inducible T7 RNA polymerase/promoter double plasmid expression vectors pGP1-2 and pT7-7 [Tabor, S., & Richardson, C. C. (1985) Proc. Natl. Acad. Sci. U.S.A. 82, 1074-1078]. The pig citrate synthase cDNA was modified to delete the N-terminal leader sequence; then by use of a synthetic oligonucleotide linker, the modified cDNA was cloned into pT7-7 immediately following the initiator Met. A glutamate-requiring (citrate synthase deficient), recA- E. coli mutant, DEK15, was transformed with pGP1-2 and then pT7-7PCS. pT7-7PCS complemented the E. coli gltA mutation.(ABSTRACT TRUNCATED AT 250 WORDS)
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DOI：
10.1073/pnas.82.4.1074
发表时间：
1985-01-01
影响因子：
11.1
作者：
TABOR, S;RICHARDSON, CC
通讯作者：
RICHARDSON, CC
DOI：
10.1016/s0021-9258(18)48074-x
发表时间：
1987-07
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
S. Karnik;M. Nassal;T. Doi;E. Jay;V. Sgaramella;H. Khorana
通讯作者：
S. Karnik;M. Nassal;T. Doi;E. Jay;V. Sgaramella;H. Khorana
哺乳动物线粒体在体外导入和加工杂合蛋白。
DOI：
--
发表时间：
1986
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Nguyen,M;Argan,C;Lusty,CJ;Shore,GC
通讯作者：
Shore,GC
影响因子：
56.9
作者：
HORWICH, AL;FENTON, WA;ROSENBERG, LE
通讯作者：
ROSENBERG, LE
G 蛋白 cDNA 在大肠杆菌中的表达。
DOI：
--
发表时间：
1987
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Graziano,MP;Casey,PJ;Gilman,AG
通讯作者：
Gilman,AG