SUBKLONING GEN ANTIGEN TUBERKULOSIS 85B DENGAN MENGGUNAKAN SIGNAL PEPTIDE AQ1 ENDOXILANASE

Reny Guspratiwi, Is Helianti, Abinawanto Abinawanto, Astutiati Nurhasanah

Sari


Penanggulangan penyakit Tuberkulosis dengan lebih efektif saat ini dilakukan melalui pengembangan vaksin kombinasi  yang merupakan kombinasi dari vaksin BCG dengan protein antigen yang berasal dari Mycobacterium tuberculosis. Salah satu protein antigen yang tengah diteliti adalah ag85B. Pada penelitian ini telah dilakukan kloning gen antigen Tuberkulosis ag85B di Eschericia coli yang digabungkan dengan signal peptid dari gen Bacillus subtilis AQ1 endoxilanase. Gen antigen 85B yang berukuran 991 pb diklona ke dalam vektor pUC57 yang mengandung promoter dan signal peptid AQ1 endoxilanase (pUC57 fragmen 2).  Vektor pUC57 fragmen 2 berukuran sekitar 2500 pb. Oleh karena itu menghasilkan plasmid rekombinan yang berukuran sekitar 3500 pb. Hasil ekspresi dari plasmid rekombinan diuji dengan SDS-PAGE.  Hasil penelitian menunjukkan gen antigen 85B berhasil diklona ke dalam vektor pUC57 fragmen 2 akan tetapi uji ekspresi dengan SDS-PAGE menunjukkan gen ag85B tersebut belum dapat diekspresikan

 

More effective prevention of Tuberculosis is now done through the development of vaccines combination, which is combined BCG vaccine with antigen from Mycobacterium tuberculosis. One of the antigen proteins under investigation is Ag85B. Cloning of Tuberculosis antigen Ag85B with signal peptide AQ1 endoxylanase from Bacillus subtilis gene in Escherichia coli DH5α was done in this research. Antigen 85B’s gene sized 991 bp was cloned into 2500 bp of pUC57 vector containing promoter and signal peptide AQ1 endoxylanase (fragment 2 of pUC57). It therefore produced a recombinant plasmid around 3500 bp. The expression of recombinant plasmid was tested by SDS-PAGE. The Result of this research showed that antigen 85B’s gene was successfully cloned into fragment 2 of pUC57 vector, but the expression was tested by SDS-PAGE showed that antigen 85B’s gene has not yet been expressed.

 

Keyword          : Antigen 85B, E. coli DH5 α, pUC57 fragmen2, signal peptide


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