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首页> 外文期刊>Life sciences >HETEROLOGOUS EXPRESSION OF CARBONYL REDUCTASE - DEMONSTRATION OF PROSTAGLANDIN 9-KETOREDUCTASE ACTIVITY AND PARAQUAT RESISTANCE
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HETEROLOGOUS EXPRESSION OF CARBONYL REDUCTASE - DEMONSTRATION OF PROSTAGLANDIN 9-KETOREDUCTASE ACTIVITY AND PARAQUAT RESISTANCE

机译：羰基还原酶的异表达-前列腺素9-酮还原酶活性和百草枯抗性的证明。

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摘要

Transfection of murine NIH3T3 fibroblasts with a pSV2-derived eukaryotic expression vector for human cytosolic carbonyl reductase (E.C. 1.1.1.141) resulted in clones with increased carbonyl reductase activity as demonstrated by an elevation in cellular NADPH-dependent alcohol (menadione) reductase activity. Prostaglandin 9-ketoreductase (9KR) activity, previously noted only in purified enzyme preparations, was also elevated. Although the cellular molar capacity of 9KR activity was less than menadione reductase activity (picomoles versus nanomoles per mg of protein), when compared to endogenous activity there was a greater relative increase in 9KR activity as compared to menadione activity (10 fold increase versus 3 fold). Thus, the 9KR properties of carbonyl reductase may have a physiologic role in prostaglandin regulation. Most transgenic clones lost their enhanced carbonyl reductase activity despite continuous selection, but two clones retained enhanced enzyme activity. RNA analysis indicated that these two murine clones expressed human carbonyl reductase mRNA. These two clones overexpressing carbonyl reductase did not display resistance to menadione, in agreement with a previous report. There was, however, a demonstrable increase in resistance to paraquat of a magnitude similar to that previously noted with transgenic cell lines overexpressing manganese superoxide dismutase. [References: 30]
机译：用pSV2衍生的人胞质羰基还原酶（E.C. 1.1.1.141）真核表达载体转染鼠NIH3T3成纤维细胞后，克隆的羰基还原酶活性增强，细胞NADPH依赖性醇（甲萘醌）还原酶活性升高。以前仅在纯化的酶制剂中注意到的前列腺素9-酮还原酶（9KR）活性也提高了。尽管9KR活性的细胞摩尔容量小于甲萘醌还原酶活性（皮摩尔对纳摩尔/毫克蛋白质），但与内源性活性相比，9KR活性相对于甲萘醌活性有更大的相对增加（增加10倍对3倍） ）。因此，羰基还原酶的9KR性质可能在前列腺素调节中具有生理作用。尽管连续选择，大多数转基因克隆仍丧失了增强的羰基还原酶活性，但是两个克隆保留了增强的酶活性。 RNA分析表明，这两个鼠类克隆表达了人类羰基还原酶mRNA。与先前的报道一致，这两个过表达羰基还原酶的克隆均未显示出对甲萘醌的抗性。然而，对百草枯的抗性明显增加，其幅度与先前过表达锰超氧化物歧化酶的转基因细胞系相似。 [参考：30]

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