The Good Conditions Exploring For Secretion Of Haps Protein Of Non-typeable Haemophilus Influenzae And Hap_s Purification

ObjectiveTo check the factor requirements of the non-typeable Haemophilus influenzae ATCC49247, and to explore the good conditions for the secretion of Haps protein of the non-typeable Haemophilus influenzae ATCC49247, and to purify Hap_s by weak cation exchange columns of CM Sepharose Fast Flow, and optimize the elution condition .Methods1. Smear NTHi ATCC49247 on LB plate medium homogeneous,then put three filter paper on it, dropping 20μl solution on each filter paper, one is Hemin solution, one is β-NAD solution, and the last is admixture with both Hemin and β-NAD solution.2. To culture non-typeable Haemophilus influenzae ATCC49247 with supplemented Brain - heart infusion (sBHI) medium , and prolong the culture times in the sBHI medium, then observe the both changes of absorbency of cultured liquid at 600 micrometer (A₆₀₀) and secretion of Hap_s in the supernatants.