Kostichka K, Tao L, Bramucci M, Tomb J-F, Nagarajan V, Cheng Q
Biological and Chemical Sciences and Engineering, Central Research and Development, Experimental Station E328/B48, E. I. DuPont de Nemours Inc., Wilmington, DE 19880-0328, USA.
Appl Microbiol Biotechnol. 2003 Jul;62(1):61-8. doi: 10.1007/s00253-003-1242-6. Epub 2003 Mar 11.
Exploration of metabolically diverse rhodococci is generally hampered by the lack of genetic tools. A small cryptic plasmid (pAN12) isolated from Rhodococcus erythropolis strain AN12 was sequenced. Plasmid pAN12 encodes proteins that share homology to replication proteins and putative cell division proteins. Based on in vitro transposon mutagenesis, we determined that the Rep protein of pAN12 is essential for plasmid replication in Rhodococcus spp., and the putative cell division protein Div is important for plasmid stability. The pAN12 replicon is able to replicate in R. erythropolis strains AN12 and CW23 (ATCC 47072) and is compatible with the nocardiophage Q4 replicon present on a Rhodococcus shuttle plasmid pDA71. pAN12 appears to belong to the pIJ101/pJV1 family of rolling circle replication plasmids. Expression of an isoprenoid pathway gene ( dxs) on the pAN12-derived multicopy shuttle vector increased production of carotenoid pigments in R. erythropolis ATCC 47072.
缺乏遗传工具通常会阻碍对代谢多样的红球菌的研究。对从红平红球菌菌株AN12中分离出的一个小的隐蔽质粒（pAN12）进行了测序。质粒pAN12编码与复制蛋白和假定的细胞分裂蛋白具有同源性的蛋白质。基于体外转座子诱变，我们确定pAN12的Rep蛋白对红球菌属中质粒的复制至关重要，而假定的细胞分裂蛋白Div对质粒稳定性很重要。pAN12复制子能够在红平红球菌菌株AN12和CW23（ATCC 47072）中复制，并且与存在于红球菌穿梭质粒pDA71上的诺卡氏噬菌体Q4复制子兼容。pAN12似乎属于滚环复制质粒的pIJ101/pJV1家族。pAN12衍生的多拷贝穿梭载体上类异戊二烯途径基因（dxs）的表达增加了红平红球菌ATCC 47072中类胡萝卜素色素的产量。
