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[Construction of expression vectors for alkaliphilic Bacillus sp. N16-5]
Objective:
To construct an expression vector for alkaliphilic Bacillus sp. N16-5.
Methods:
Bacillus subtilis expression vector pHCMC04 was used as a backbone. Its xylose-inducible promoter cassette was replaced by the constitutive promoters P43 (from B. subtilis) and P(EF) (from Bacillus sp. N165), separately, resulting in two expression vectors pABN165P43 and pABN165P(EF). Green fluorescent protein gene gfp was linked to the two vectors as a reporter gene. Fluorescence microscope and multifunctional fluorescent reader were used to test the expression efficiency of the system.
Results:
Green fluorescence was visualized in Bacillus sp. N16-5 with pABN165P(EF)-gfp or pABN165P43-gfp. Quantitative data analysis revealed that fluorescence was first detected around the 7th hour. The fluorescence intensity increased rapidly from the 7th hour to the 12th hour and reached the maximum at about the 12th hour.
Conclusion:
Two expression vectors for Bacillus sp. N16-5 have been constructed, allowing expression of exogenous protein in alkaliphilic Bacillus sp. N165.
