Isolation of the yeast structural gene for the membrane-associated enzyme phosphatidylserine synthase.
Isolation of the yeast structural gene for the membrane-associated enzyme phosphatidylserine synthase.
复制标题
膜相关酶磷脂酰丝氨酸合酶的酵母结构基因的分离。
DOI：
10.1073/pnas.80.23.7279
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发表时间：
1983
影响因子：
11.1
通讯作者：
Henry,SA
中科院分区：
文献类型：
--
作者：
Letts,VA;Klig,LS;Bae-Lee,M;Carman,GM;Henry,SA
磷脂酰丝氨酸合成酶（CDP二酰基甘油：L-丝氨酸O-磷脂酰转移酶，EC 2.7.8.8）的结构基因（CHO 1）通过在酿酒酵母中的遗传互补从嵌合质粒上的酵母基因组DNA库中分离。克隆的DNA（4.0个内切酶长）显示代表酵母基因组中的独特序列。整合质粒上的DNA序列显示重组到CHO 1基因座中，确认其遗传同一性。在自主复制质粒上转化该基因的cho 1酵母菌株显着增加了受调节的膜相关酶磷脂酰丝氨酸合酶的活性。从该转化菌株的微粒体中部分纯化磷脂酰丝氨酸合成酶证实，与野生型菌株相比，该膜结合酶过量产生6至7倍。该菌株还合成产物磷脂，磷脂酰丝氨酸，在增加的速度。转化的菌株改变了各种其他磷脂的比例，这表明它们的合成受到酵母中磷脂酰丝氨酸合成速率的影响。
The structural gene (CHO1) for phosphatidylserine synthase (CDPdiacylglycerol:L-serine O-phosphatidyltransferase, EC 2.7.8.8) was isolated by genetic complementation in Saccharomyces cerevisiae from a bank of yeast genomic DNA on a chimeric plasmid. The cloned DNA (4.0 kilobases long) was shown to represent a unique sequence in the yeast genome. The DNA sequence on an integrative plasmid was shown to recombine into the CHO1 locus, confirming its genetic identity. The cho1 yeast strain transformed with this gene on an autonomously replicating plasmid had significantly increased activity of the regulated membrane-associated enzyme phosphatidylserine synthase. Partial purification of phosphatidylserine synthase from microsomes of this transformed strain confirmed that the membrane-bound enzyme was overproduced 6- to 7-fold as compared with the wild-type strain. The strain also synthesized the product phospholipid, phosphatidylserine, at an increased rate. The transformed strain had altered proportions of a variety of other phospholipids, suggesting that their synthesis is affected by the rate of synthesis of phosphatidylserine in yeast.