PEMBUATAN ANTIBODI REKOMBINAN BEGOMOVIRUS ISOLAT CABAI
| Main Authors: | , Yashanti Berlinda Paradisa, , Dr.Ir. Sri Sulandari, S.U. |
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| Format: | Thesis NonPeerReviewed |
| Terbitan: |
[Yogyakarta] : Universitas Gadjah Mada
, 2012
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| Subjects: | |
| Online Access: |
https://repository.ugm.ac.id/99069/ http://etd.ugm.ac.id/index.php?mod=penelitian_detail&sub=PenelitianDetail&act=view&typ=html&buku_id=55493 |
Daftar Isi:
- Pepper yellow leaf curl disease caused by begomovirus is one of Indonesia's pepper production constraints. The disease is widespread and very difficult to control. One way to prevent such diseases by early detection through serological test. The main obstacle for begomovirus serological test is the difficulty in preparing of pure viruses to make antibodies. Through cloning of begomovirus envelope protein-coding genes have been produced recombinant proteins that could be used as an immunogen to create recombinant antibodies. This study aimed to produce of recombinant antibodies that could be used as an early detection of Begomovirus accurately, faster, and cheaper to prevent that disease epidemics. The study was conducted by culturing a recombinant plasmid PepYLCIV isolates Congkrang. Recombinant plasmid was verified by PCR analysis and DNA sequencing. Recombinant plasmids was cultured in liquid LB medium containing ampicillin 50 ug / ml and induced by IPTG until final concentrations became 1mM to obtain the recombinant protein was then used as antigen. Recombinant protein was isolated and characterized by SDS-PAGE to determine the molecular weight and is used as the immunogen. Immunization of rabbits with a dose of 400 Î1⁄4g and the injection of 1-week interval. The resulting antiserum was purified using the method of almonium sulfate saturation. Purity of the recombinant antibodies were analyzed using a spectrophotometer. Titer testing done using the I-ELISA. The resulting antibodies are used to test the pepper plants and weeds Ageratum conyzoides infected using the I-ELISA and DIBA. The results of verification by PCR and DNA sequence analysis showed that the recombinant plasmid still contains the recombinant of DNA PepYLCIV. Expression of coat protein genes a PepYLCIV isolates Congkrang produce a recombinant protein bands measuring approximately 29kDa. Crude antisera and purified antibodies produced has a concentration of respectively 0.389 mg / ml and 0.227 mg / ml. Through the I-ELISA and DIBA test, crude antiserum could be used to detect the presence of begomovirus in pepper plants and weeds Ageratum conyzoides symptomatic yellow curls.