Introduction   :

Unmet need for contraception affect as many as 818 million sexually productive women in developing countries.The most cited reasons are fear of side effects, beliefs, and ignorance. A novel approach is needed to tackle above reasons.

Bacterial Contraception is a novel contraceptive method using engineered normal vaginal microflora. Microflora is engineered to secrete spermicidal peptide, subtilosin A, that can be controlled with genetic toggle switch. This pilot study aims to measure the ability of E. coli BL21 producing subtilosin A, assess its biological activity, and assess the capability of genetic toggle switch.

Method           :

BL21 is engineered to produce subtilosin A by inserting sboA gene to pET9a plasmid by restriction-ligation at NdeI and BamHI. SDS-PAGE is used to confirm peptide expression. Sperm motility and well diffusion assay against M.luteus are used to assess biological activity of subtilosin A produced by BL21.

Qualitative analysis is used for genetic toggle switch assessment. LB agar infused with IPTG and xylose are used to test fluorescence protein expression.

Result             :

SDS-PAGE analysis confirmed subtilosin A at 3kDa range.Sperm motility assay shows motility reduction in subtilosin A producing group without significant result compared with control group.Antimicrobial activity to M.luteus shows no significant activity.

Genetic toggle switch activity confirmed qualitatively by observing green light fluorescence. Red light fluorescence can not be observed.

Conclusion     :

This pilot study show that BaCon could produce subtilosin A. More works from multiple fields are needed to make it functional.