THE PRODUCTION OF LACTOFERRIN-PRODUCING BACTEROIDES UNIFORMIS

ABSTRACT

Abstract To construct lactoferin-producing Bacteroides uniformis, the human lactoferrin gene(hLF) was inserted into pVAL-1, a E. coli-Bacteroides shuttle vector, to generate the recombinantplasmid pVLFK. Then, withE. coli HB101 (R751) as a donor either pVAL-1 or pVLFKwas transferred intoB. uniformis BU1001. The erythromycin resistant strains were selected andisolated as a plasmid containing strain. By PCR technique, the lactoferrin gene was detected inthe bacteria cell extract from each strain. The presence ofhLF has been shown as a 2.1 Kbamplicon inB. uniformis strain pVLFK. The expression of hLF was assessed by Northernhybridization and the gene product was detected by Western blot analysis with anti-humanlactoferrin monoclonal antibody. Although the transcription level ofhLF gene was extremelylow in the strain harboring pVLFK, the specific protein was detected by Western blot analysis.However the size of gene product in this strain was higher than that of standard human LF(SIGMA). The results revealed that a lactoferrin gene bearing plasmid could replicate and wasexpressed inB. uniformis strain pVLFK. The properties of this strain will be further study.Chiang Mai Med Bull 2001;40(4)187-194.