Long-chain fatty acid transport in Escherichia coli. Cloning, mapping, and expression of the fadL gene.
Long-chain fatty acid transport in Escherichia coli. Cloning, mapping, and expression of the fadL gene.
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DOI：
10.1016/s0021-9258(18)89661-2
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发表时间：
1985-02
期刊：
影响因子：
--
通讯作者：
P. Black;S. Kianian;C. DiRusso;W. D. Nunn
中科院分区：
文献类型：
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作者：
P. Black;S. Kianian;C. DiRusso;W. D. Nunn
长链脂肪酸穿过大肠杆菌K-12的内膜的转运需要功能性fadL基因（Maloy，S. R.，金斯伯格角L.，西蒙斯河W.，和纳恩，W. D.（1981）J.Biol.Chem.256，3735-3742）。用二维pI/十二烷基硫酸钠（SDS）聚丙烯酰胺凝胶电泳（Ginsburgh，C. L.，布莱克，P. N.，和纳恩，W. D.（1984）J.Biol.Chem.259，8437-8443）。为了确定fadL基因是否是这种33，000-道尔顿蛋白的结构基因，我们克隆、定位并分析了fadL基因的表达。fadL基因定位于大肠杆菌2.8-脱氢酶EcoRV片段上。coli基因组DNA。含有该基因的疟原虫（i）补充所有的fadL突变体，（ii）使携带它们的fadL菌株的长链脂肪酸转运活性增加2- 3倍，和（iii）指导具有与Ginsburgh等所述相同分子量和等电点的膜蛋白的合成。这是一种表观分子量为43，当在SDS存在下于100 ° C溶解时为33，000道尔顿，当在SDS存在下于50 ° C溶解时为33，000道尔顿。
Transport of long-chain fatty acids across the inner membrane of Escherichia coli K-12 requires a functional fadL gene (Maloy, S. R., Ginsburgh, C. L., Simons, R. W., and Nunn, W. D. (1981) J. Biol. Chem. 256, 3735-3742). Mutants defective in the fadL gene lack a 33,000-dalton inner membrane protein as evaluated using two-dimensional pI/sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis (Ginsburgh, C. L., Black, P. N., and Nunn, W. D. (1984) J. Biol. Chem. 259, 8437-8443). In an effort to determine whether the fadL gene is the structural gene for this 33,000-dalton protein, we have cloned, mapped, and analyzed the expression of the fadL gene. The fadL gene has been localized on a 2.8-kilobase EcoRV fragment of E. coli genomic DNA. Plasmids containing this gene (i) complement all fadL mutants, (ii) increase the long-chain fatty acid transport activity of fadL strains harboring them by 2- to 3-fold, and (iii) direct the synthesis of a membrane protein which has the same molecular weight and isoelectric point as that described by Ginsburgh et al. This is a heat-modifiable protein which has an apparent molecular weight of 43,000 daltons when solubilized at 100 degrees C in the presence of SDS and 33,000 daltons when solubilized at 50 degrees C in the presence of SDS.