Judul: Uji Kemampuan Kandidat Vaksin DNA Viral Nervous Necrosis Dalam Menginduksi Antibodi Pada Ikan Kerapu Tikus (Cromileptes altivelis)
Abstrak: Viral nervous necrosis (VNN) is one of the diseases that often infect grouper fish. Two DNA vaccines for VNN, namely pmBA-CP and pJfKer-CP have been constructed. The pmBA- CP construct is controlled by medaka β-actin (mBA) promoter, while pJfKer-CP by Japanese flounder keratin (JfKer) promoter. This study was aimed to examine the antibody induction of coat protein (CP) DNA vaccinated humpback grouper (Cromileptes altivelis). The research was divided into three steps, i.e. (1) analysis of the CP VNN using SDS-PAGE and Western blot methods, (2) analysis of CP mRNA expression using reverse transcriptase PCR (RT-PCR) method, and (3) analysis of anti VNN antibody induction of vaccinated fish using enzyme linked immunosorbent assay (ELISA) sandwich method, and VNN challenge test of vaccinated fish. The results of SDSPAGE showed a specific protein of about 42 kDa from DNA vaccine-injected catfish embryos. By Western blot analysis using a polyclonal antibody produced in this study, the similar size of protein was also obtained in catfish embryos injected by DNA vaccine. This suggests that the polyclonal antibody anti VNN from rabbit immunization with VNN could recognize CP protein from DNA vaccine injected catfish embryos. In subsequent experiment, we used pmBA-CP DNA construct due to the result of CP protein from pmBA-CP injected catfish embryos was higher than that of pJfKer-CP. Humpback grouper juvenile was intramuscularly injected with pmBA-CP in a dose of 12.5 μg per fish, and RTPCR analysis were performed to detect the present of CP mRNA in the fish muscle extracted at different time post vaccination. The results showed that CP mRNA expression was detected at 24 hours postvaccination. This means that mBA promoter could regulate the expression of CP gene in humpback grouper. Furthermore, the result of ELISA showed higher ratio of S/P using serum from pmBA-CP vaccinated fish than unvaccinated fish (control group) at 21 to 35 days postvaccination. This suggests that DNA vaccination is capable of inducing the antibody anti VNN in humpback grouper. Furthermore, the result of challenge test with VNN virus titer of 103,5 FID50/0,2 ml generated 60% relative percent survival. Thus, pmBA-CP DNA vaccination can be useful to improve the resistance of grouper fish against VNN infection, and support the increasing aquaculture production level.
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Judul: Development of DNA Vaccine Encoding Glycoprotein of Koi Herpes Virus Using Local Isolate.
Abstrak: The aim of this research was to develop DNA vaccine of Koi Herpesvirus in carp. Glycoprotein gene originated from ORF 25 KHV virus that is 1.8 kbp in size, is a potential gene in DNA vaccine construction. The sequencing and similarity analysis of the gene using BLAST software showed that this gene is a GP25 gene and was found to be very similar (99%) to GP25 of KHV virus from the USA and Israel. When this GP25 gene was ligated with pGEMT Easy (pT, 3 kbp in size) vector, plasmid size was increasing up to 5 kb. Following this, a digestion of pT-GP25 plasmid using Sal I enzyme was applied resulted in a GP25 fragment (1.8 kbp). Subsequently, pAct-D6 with a size of 8.5 kbp was also cut using the same enzyme to eleminate a D6 gene with a size of 1.6 kbp. GP25 gene was then ligated with pAct resulted a pAct-GP25 plasmid with a size of 8.8 kbp. As there was only one restriction site applied, ligation can be occurred in two different directions. Thus, in order to determine the plasmid with the desired direction PCR analysis was carried out, and plasmid with the right direction was the one that sized approximately 2.1 kbp (no. 6, 17 and 20). High level of GFP gene expression was detected in all tissues analyzed, i.e. kidney, gil, spleen, and muscle 24 hours after injection which indicated that β-actin promoter of Japanese medaka could be active in common carp. Although not as high as on 24 hours, expression was still detected until day 7 and disappeared after day 28 post injection indicating that pAct-GFP can be remained and β-actin promoter of Japanese medaka can be active in muscle, gills and kidney at least a week after injection. The result of RT-PCR analysis on pAct-GP25 injected common carp showed that GP25 gene expression can be detected 14 days after injection in all tissues observed. The expression level differences between GFP and GP25 genes possibly related to the primer sensitivity on annealing process to cDNA template and the size of PCR targeted DNA fragment, as smaller PCR targeted product will be easier to be amplified than a bigger one. The length of PCR targeted DNA with GP25 primer was 1.8 kbp whereas GFP gene was 0.6 kbp.The challenge test on common carp results showed that all unvaccinated and negative control (not infected with KHV) was alive until the end of the experiment. Whereas, the survival of fish vaccinated with a dose of 12.5 was ranged from 60.9 and 96.7%. In relation to the activity of β-actin promoter that was used in this DNA vaccine, gene expression was occured 24 hours after vaccination and remained until 2 weeks. The control fish that was not injected by GP25 showed no GP25 expression either in the observed tissues or organs and resulted in a lower survival, i.e. 23.3%. The challenge test results has indicated that the fish vaccinated with GP25 at doses of 7.5 and 12.5 μg/100μL were able to produce KHV glycoprotein in their body which would be
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Judul: Perancangan jalur interpretasi wisata sejarah kawasan kota tua Jakarta
Abstrak: Tapak perancangan berupa jalur interpretasi yang terletak di dalam wilayah administrasi kota Jakarta Barat dan melalui Jl. Stasiun, Jl. Pintu Besar Selatan dan Utara, Jl. Kali Besar timur dan barat, Jl. Cengkeh, Jl. Pos, dan Jl. Lada. Aksesibilitas menuju tapak mudah dengan berbagai alternatif sarana transportasi. Tata guna lahan kawasan 78% terdiri dari perkantoran perdagangan.
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