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Peningkatan resistensi udang windu penaeus monodon terhadap penyakit white spot syndrome virus melalui transfer gen penaeus monodon antiviral
| Content Provider | Semantic Scholar |
|---|---|
| Author | Parenrengi, Andi |
| Copyright Year | 2010 |
| Abstract | ANDI PARENRENGI. The Improvement of Tiger Shrimp Diseases Resistance against White Spot Syndrome Virus by Transfer of Penaeus monodon Antiviral Gene. Under direction of KOMAR SUMANTADINATA, ALIMUDDIN, and SUKENDA. Shrimp is one of the most important species in aquaculture. Tiger shrimp Penaeus monodon, an indigenous crustacean species in Indonesia, has been widely reared in brackish water pond. During the last decade, the worldwide shrimp culture was greatly puzzled by diseases caused by viruses and suffered significant economic losses. White spot syndrome virus (WSSV) is the most threatening infectious agent in shrimp aquaculture. Production of diseases-resistant shrimp by genetic manipulation is an alternative way to elucidate the problem. Therefore, the aim of this study was to improve the shrimp resistance to WSSV by introducing an antiviral gene isolated from tiger shrimp P. monodon. ProAV promoter and cDNA of PmAV antiviral gene were isolated from tiger shrimp using PCR and RT-PCR methods, respectively. The results showed the success in isolating the tiger shrimp ProAV promoter sequence of 368 bp in length. BLAST-N analysis showed that the promoter has high similarity (95-98%) compared with the other promoters presented in the GeneBank. The existence of the important transcription factors in promoter regulation such as TATA box, MRE, TCF-1, SP-1, GAL-4, and GATA-1 were identified in the promoter. The PmAV antiviral cDNA of 513 bp in length that consists of 170 amino acid residues has also been successfully isolated from tiger shrimp. BLAST-N analysis showed the high similarity (100%) compared with the other antiviral genes deposited at the GeneBank. Amino acid deduction analysis (BLAST-P) of PmAV cDNA revealed a C-type lectin-like domain (CTLD) that is similar with the C-type lectin gene isolated from several crustacean species. Furthermore, the isolated ProAV and PmAV sequences were ligated to pEGFP-N1 and pBluescript-SK, respectively to construct pProAV-EGFP and pProAV-PmAV expression vectors. Expression vectors were transferred into embryos by transfection method using JetPEI reagent. ProAV promoter activity was determined by analyzing EGFP gene expression as a reporter using RT-PCR method. The result showed that ProAV promoter was able to drive EGFP gene expression on embryo and larvae of tiger shrimp. Transient EGFP expression analysis showed a similar pattern with the PmAV gene expression, where they started to express at 12 hours after transfection (hat), the peak expression level at 24 hat, and then decreased slightly at 30 hat. Thus, pProAV-PmAV was then constructed, and F0 transgenic shrimp was produced to examine the PmAV activity against WSSV infection. Result of challenge test showed that PmAV gene expression in F0 transgenic PL-25 was upregulated during WSSV infection, and over-expression of PmAV cDNA increased the survival rate of 24.5% in transgenic shrimp (95.6% survived) compared to the control shrimp (71.1% survived) against WSSV. The body weight and length of 1.5 months transgenic shrimp did not show significantly difference with the non-transgenic shrimp. Therefore, the use of transgenic shrimp over-expressing PmAV cDNA may be helpful to recovery Indonesian shrimp aquaculture. |
| File Format | PDF HTM / HTML |
| Alternate Webpage(s) | https://repository.ipb.ac.id/bitstream/handle/123456789/55056/2010apa.pdf?isAllowed=y&sequence=3 |
| Alternate Webpage(s) | https://repository.ipb.ac.id/bitstream/handle/123456789/55056/COVER.pdf?isAllowed=y&sequence=12 |
| Language | English |
| Access Restriction | Open |
| Content Type | Text |
| Resource Type | Article |
