Respon imun mencit BALB/c yang diimunisasi dengan protein CA murni hasil ekspresi klon glen gag-ca virus penyakit Jembrana

SOYI, Dwi Susilowati, Dr.drh. Asmarani Kusumawati, MP

2006 | Tesis | S2 Bioteknologi

Penyakit Jembrana merupakan penyakit akut yang menginfeksi sapi Bali. Penyakit Jembrana ini disebabkan oleh suatu Lentivirus, yang merupakan anggota famili Retroviridae dan subfamili Lentiviridae, virus ini dinamakan virus penyebab penyakit Jembrana (VPJ) atau Jembrana disease virus (JDV). Calon vaksin protein untuk penyakit Jembrana, dikonstruksi dengan menyisipkan gen gag-ca yang menyandi protein capsid (CA) ke dalam vektor ekspresi prokariot ialah pGEX-2T, sehingga menghasilkan plasmid pGEX-CA. Penelitian ini bertujuan untuk memperoleh protein CA murni dari hasil ekspresi klon gen gagca melalui purifikasi dengan kolom afinitas dan pemotongan dengan Thrombin Protease serta mengetahui respon imun humoral mencit BALB/c yang ditimbulkan oleh imunisasi dengan protein CA. Hasil penelitian menunjukkan bahwa protein CA murni dapat diperoleh melalui purifikasi dengan kolom afinitas menggunakan MicroSpin GST dan pemotongan dengan Thrombin Protease. Imunisasi protein CA pada mencit BALB/c memberikan respon imun humoral yang masih rendah berupa produksi antibodi anti-CA.

Jembrana disease is an acute disease which attack Bali cattle. The disease was caused by a Lentivirus, a member of Retroviridae family and subfamily Lentiviridae, which was called as Jembrana Disease Virus (JDV). Protein vaccine candidate for the disease constructed by inserting gag-ca gene that encodes capsid protein (CA) to a prokaryotic expression vector, pGEX-2T. The aim of the research is: (1) to get pure CA protein from clone expression of gag-ca gene through purification with affinity column and cleavage with protease thrombin; (2) to study humoral immune response BALB/c mice that induced by capsid protein immunization. To construct the protein vaccines was examine by these procedures: pGEX-CA plasmid isolation, PCR, recombinant protein isolation, cleavage of fusion protein by using protease thrombin, and immunization to BALB/c mice. The immune response was measured by indirect ELISA technique to BALB/c mice serum. Which it was injected intraperitonially by the capsid protein with the dosage of 5 ul at 0, 3, and 6 weeks. Serums were collected at 2 weeks after the last vaccination. The result showed that pure capsid protein can be obtained through purification with affinity column using MicroSpin GST and cleavage using protease thrombin. Capsid protein immunization on BALB/c mice induce low humoral immune response that produced antibodies against CA antigen.

Kata Kunci : Vaksin Protein CA,Gen gag,ca,JDV,Jembrana disease virus (JDV), protein vaccine, gag-ca gene, protein purification, humoral immune response