Lactobacillus rhamnosus is found in the human gastrointestinal tract and is important for probiotics. We became interested in L. rhamnosus isolate ATCC 8530 in relation to beer spoilage and hops resistance. We report here the genome sequence of this isolate, along with a brief comparison to other available L. rhamnosus genome sequences. Lactobacillus rhamnosus is an organism of interest for human health and probiotics. In our case, we became interested in a particular L. rhamnosus isolate due to its ability to spoil beer. Among five L. rhamnosus isolates tested, only one (ATCC 8530) was able to grow in beer (1). We therefore performed further analyses on this isolate and found that it was among a few lactic acid bacteria (LAB) that showed increased resistance to hops in the presence of ethanol (2). This finding was particularly intriguing considering the antimicrobial properties of both compounds and because hops has been suggested as a replacement for antibiotics to control LAB contamination in the fuel ethanol industry (6). To get a better understanding of how L. rhamnosus ATCC 8530 grows in beer and resists hops, we sequenced its genome for comparative analysis with other sequenced LAB.Sequencing was done using the Roche 454 Genome Sequence FLX platform. Paired and unpaired reads were obtained during two different runs, giving a final coverage of ϳ30ϫ. A total of 520,000 reads were assembled using Newbler GS De Novo assembler, giving 34 contigs split among six scaffolds. The scaffolds were then ordered according to the L. rhamnosus Lc 705 genome, and gaps were closed by sequencing PCR amplicons via the ABI 3700xl platform. The final circularized genome is 2,960,339 bp with a GϩC content of 46.8%.Genome annotation was done via the Institute for Genome Sciences Annotation Engine, which predicted 2,904 coding sequences (CDS), 60 tRNA, and five rRNA operons. Of the predicted CDS, ϳ700 are hypothetical proteins with unknown function. Unlike L. rhamnosus Lc 705 and HN001, no plasmids are found in this isolate.Comparative analysis with L. rhamnosus ATCC 8530 and the four other currently available L. rhamnosus genome sequences (3, 4) showed that L. rhamnosus ATCC 8530 is least similar to the probiotic isolates GG and HN001. In contrast, the intestinal isolate L. rhamnosus LMS2-1 is very similar to ATCC 8530, with most (Ͼ90%) predicted proteins having Ն99% identity. Preliminary analyses indicate that only four regions are different between L. rhamnosus ATCC 8530 and LMS2-1, all related to mobile elements (L. rhamnosus ATCC 8530 has no predicted bacteriophagerelated genes). The majority of L. rhamnosus ATCC 8530 was also conserved with the dairy industry isolate L. rhamnosus Lc 705, except for one distinct location missing in the former isolate and several bacteriophage-related regions not found in the latter isolate, respectively.Considering the high degree of similarity between L. rhamnosus ATCC 8530 and LMS2-1, further analyses will be done to determine if the latter isolate can grow in beer, as well as to show a co...