Poultry Microbiological Safety Research Unit, Richard B. Russell Agricultural Research Center, Agricultural Research Service, United States Department of Agriculture, Athens, GA 30605, USA.
Plasmid. 2013 Sep;70(2):268-71. doi: 10.1016/j.plasmid.2013.04.004. Epub 2013 Apr 30.
The complete nucleotide sequence was determined for a cryptic plasmid, pTIW94, recovered from several Campylobacter jejuni isolates from wild birds in the southeastern United States. pTIW94 is a circular molecule of 3860 nucleotides, with a G+C content (31.0%) similar to that of many Campylobacter spp. genomes. A typical origin of replication, with iteron sequences, was identified upstream of DNA sequences that demonstrated similarity to replication initiation proteins. A total of five open reading frames (ORFs) were identified; two of the five ORFs demonstrated significant similarity to plasmid pCC2228-2 found within Campylobacter coli. These two ORFs were similar to essential replication proteins RepA (100%; 26/26 aa identity) and RepB (95%; 327/346 aa identity). A third identified ORF demonstrated significant similarity (99%; 421/424 aa identity) to the MOB protein from C. coli 67-8, originally recovered from swine. The other two identified ORFs were either similar to hypothetical proteins from other Campylobacter spp., or exhibited no significant similarity to any DNA or protein sequence in the GenBank database. Promoter regions (-35 and -10 signal sites), ribosomal binding sites upstream of ORFs, and stem-loop structures were also identified within the plasmid. These results demonstrate that pTIW94 represents a previously un-reported small cryptic plasmid with unique sequences as well as highly similar sequences to other small plasmids found within Campylobacter spp., and that this cryptic plasmid is present among Campylobacter spp. recovered from different genera of wild birds.
从美国东南部野生鸟类中分离的空肠弯曲菌分离株中回收了一个隐秘质粒 pTIW94,其完整的核苷酸序列已被确定。pTIW94 是一个 3860 个核苷酸的圆形分子,其 G+C 含量(31.0%)与许多弯曲菌属的基因组相似。在与复制起始蛋白具有相似性的 DNA 序列上游,鉴定出了一个典型的复制起点,带有回文序列。共鉴定出 5 个开放阅读框(ORF);其中 5 个 ORF 中的两个与在大肠弯曲菌中发现的质粒 pCC2228-2 具有显著相似性。这两个 ORF 与必需的复制蛋白 RepA(100%;26/26 aa 同一性)和 RepB(95%;327/346 aa 同一性)相似。鉴定出的第三个 ORF 与大肠弯曲菌 67-8 中的 MOB 蛋白(最初从猪中回收)具有显著相似性(99%;421/424 aa 同一性)。另外两个鉴定出的 ORF 要么与其他弯曲菌属的假设蛋白相似,要么与 GenBank 数据库中的任何 DNA 或蛋白质序列没有显著相似性。该质粒中还鉴定出了启动子区域(-35 和-10 信号位点)、ORF 上游的核糖体结合位点和茎环结构。这些结果表明,pTIW94 代表了一种以前未报道的具有独特序列和与弯曲菌属中其他小质粒高度相似序列的小隐秘质粒,并且这种隐秘质粒存在于从不同野生鸟类属中回收的弯曲菌属中。