title: Cloning and expression of a novel lactococcal aggregation factor from Lactococcus lactis subsp. lactis BGKP1.
abstract: BACKGROUND
  Aggregation may play a main role in the adhesion of bacteria to the gastrointestinal epithelium and their colonization ability, as well as in probiotic effects through co-aggregation with intestinal pathogens and their subsequent removal. The aggregation phenomenon in lactococci is directly associated with the sex factor and lactose plasmid co-integration event or duplication of the cell wall spanning (CWS) domain of PrtP proteinase.
  
  
  RESULTS
  Lactococcus lactis subsp. lactis BGKP1 was isolated from artisanal semi-hard homemade cheese and selected due to its strong auto-aggregation phenotype. Subsequently, non-aggregating derivative (Agg-) of BGKP1, designated as BGKP1-20, was isolated, too. Comparative analysis of cell surface proteins of BGKP1 and derivative BGKP1-20 revealed a protein of approximately 200 kDa only in the parental strain BGKP1. The gene involved in aggregation (aggL) was mapped on plasmid pKP1 (16.2 kb), cloned and expressed in homologous and heterologous lactococci and enterococci. This novel lactococcal aggregation protein was shown to be sufficient for cell aggregation in all tested hosts. In addition to the aggL gene, six more ORFs involved in replication (repB and repX), restriction and modification (hsdS), transposition (tnp) and possible interaction with mucin (mbpL) were also located on plasmid pKP1.
  
  
  CONCLUSIONS
  AggL is a new protein belonging to the collagen-binding superfamily of proteins and is sufficient for cell aggregation in lactococci.
authors.affiliation: Laboratory for Molecular Genetics of Industrial Microorganisms, Institute of Molecular Genetics and Genetic Engineering, University of Belgrade, Vojvode Stepe 444/a, P,O, Box 23, Belgrade 11010, Serbia. mkojics@eunet.rs
authors.forename: Milan
authors.initials: M
authors.lastname: Kojic
authors.forename: Branko
authors.initials: B
authors.lastname: Jovcic
authors.forename: Ivana
authors.initials: I
authors.lastname: Strahinic
authors.forename: Jelena
authors.initials: J
authors.lastname: Begovic
authors.forename: Jelena
authors.initials: J
authors.lastname: Lozo
authors.forename: Katarina
authors.initials: K
authors.lastname: Veljovic
authors.forename: Ljubisa
authors.initials: L
authors.lastname: Topisirovic
mesh_terms: D001422:Bacterial Adhesion; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D018441:Biofilms; D002611:Cheese / Q000382:microbiology; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D013294:Lactococcus lactis / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D008969:Molecular Sequence Data; D010957:Plasmids; D017422:Sequence Analysis, DNA
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial