Preparation And Charactereristics Of The Monoclonal Antibody Against MraY Protein Of Escherichia Coli

MraY, an essential membrane-embedded enzyme that catalyzes the formation of the mureinprecursor, Lipid I, from UDP-N-acetylmuramic acid-pentapeptide and the lipid carrier,undecaprenol phosphate. MraY remains one of the great hopes for developing new antibacterialsthat inhibit peptidoglycansynthesis.The bacteriophage phiX174encodes the integral membraneprotein E whose expression leads to host cell lysis by inhibition of the peptidoglycansynthesisenzyme MraY.Using target protein and assistant protein as antigen to conduct immunization in BALB/cmouse to produce monoclonal antibodies of anti-assistant and target protein, aftersubcloning forthree times, indirection ELISA and Western-Blot analysis,shows that it issuccessful to obtain1strain of hybridoma cell that cansecrete anti MraY protein monoclonal antibody(M-H11).M-H11belongs to IgG2b monoclonal antibody class and has κ light chain.To analysis thefunction of M-H11, we performed the bacterial growth inhibition test. M-H11was found to haveactivity to inhibit E.coli BL21（DE3）pLysS in vivo and vitro.Subsequently conducted aseries oftests on multidrug resistant E.coli, E.coli DH5α, E.coli DH5α（△ompT）, E.coli BL21（DE3).All found no inhabitation. M-H11retrieved its activity to inhibit E.coli when transformed theplasmid of E.coli BL21（DE3）pLysS into E.coli BL21（DE3）.