Entrapment Formulation for env-Tm Gene based on Chitosan Low Molecular Weight as a Jembrana Disease Virus Vaccine Candidate
Indra Lesmana Rahayu1, Asmarani Kusumawati2 and Ronny Martien3
1Department of Biotechnology, Postgraduate School, Gadjah Mada University, INDONESIA, 2Department of Reproduction and Obstetrics, Faculty of Veterinary Medicine, Gadjah Mada University, INDONESIA
3Department of Pharmaceutics, Faculty of Pharmacy, Gadjah Mada University, INDONESIA
Abstract
Background: Jembrana disease (JD) caused by Jembrana Disease Virus (JDV) becomes an obstacle in Bali cattle (Bos javanicus) which is local Indonesian cattle. The development of JD vaccines has an important meaning to prevent losses the Bali cattle industry in Indonesia and is one of the models in the development of the Human Immunodeficiency Virus (HIV) vaccine. Development of vaccines for JDV is carried out DNA vaccines that are expected to be able to provide better immunological effects. But, the challenge in DNA vaccines is in the delivery system of target genes to cells, so to solve this problem, research is carried out on the use of chitosan as a protective molecule as well as carrying the target gene to cells. This research was conducted at the Genetic Engineering Laboratory, Postgraduate School, Gadjah Mada University in September 2018-March 2019 Aims: The purpose of this study was to determine the low molecular weight chitosan (LC) entrapment towards pEGFP-C1-env-Tm in the formation of Chitosan Nanoparticles Low/pEGFP-C1-env-Tm complex (LC/pEGFP-C1-env-Tm). Materials and Methods: The env-Tm gene is inserted in pEGFP-C1 into the pEGFP-C1/env-Tm construct which is then transformed on the E. coli DH5α host. The construct is then formulated in the form of LC/pEGFP-C1/env-Tm complex with a low molecular weight chitosan concentration of 0.06% and the ratio of pEGFP-C1/env-Tm:LC (wt/wt) is 1:0.5-1:3. The complexes are then analyzed by gel retardation assay agarose 1%. Results: The results of this study indicate that the best entrapment results of low molecular weight chitosan to pEGFP-C1/env-Tm is in the mass ratio of pEGFP-C1/env-Tm:LC is 1:2. Conclusion: The best formulation entrapment for env-Tm by low molecular weight chitosan 0.06% is in the ratio 1:2.
Keywords: env-Tm gene, Jembrana Disease Virus, Low Molecular Weight Chitosan.
Topic: Environmentally Sustainable Agriculture