Richaud C, Higgins W, Mengin-Lecreulx D, Stragier P
J Bacteriol. 1987 Apr;169(4):1454-9. doi: 10.1128/jb.169.4.1454-1459.1987.
The Escherichia coli dapF gene was isolated from a cosmid library as a result of screening for clones overproducing diaminopimelate epimerase. Insertional mutagenesis was performed on the cloned dapF gene with a mini-Mu transposon, leading to chloramphenicol resistance. One of these insertions was transferred onto the chromosome by a double-recombination event, allowing us to obtain a dapF mutant. This mutant accumulated large amounts of LL-diaminopimelate, confirming the blockage in the step catalyzed by the dapF product, but did not require meso-diaminopimelate for growth. The dapF gene was localized in the 85-min region of the E. coli chromosome between cya and uvrD.
通过筛选过量产生二氨基庚二酸差向异构酶的克隆，从黏粒文库中分离出大肠杆菌dapF基因。用mini-Mu转座子对克隆的dapF基因进行插入诱变，产生氯霉素抗性。其中一个插入通过双重组事件转移到染色体上，从而使我们获得一个dapF突变体。该突变体积累了大量的LL-二氨基庚二酸，证实了dapF产物催化步骤中的阻断，但生长不需要内消旋二氨基庚二酸。dapF基因定位于大肠杆菌染色体85分钟区域，位于cya和uvrD之间。
