CLONING OF VP28 OPEN READING FRAME FROM WHITE SPOT SYNDROME VIRUS IN Escherichia coli DH5a
The cumulative mortality of White Spot Syndrome Virus (WSSV) infection in shrimp aquaculture may reach 100% within 10 days. VP28 is one of WSSV envelope proteins which is involved in attachment and penetration of virus into shrimp cells. The use of bacteria which expressed VP28 protein as a vaccine...
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| Format: | Final Project |
| Language: | Indonesia |
| Online Access: | https://digilib.itb.ac.id/gdl/view/10418 |
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| Institution: | Institut Teknologi Bandung |
| Language: | Indonesia |
| Summary: | The cumulative mortality of White Spot Syndrome Virus (WSSV) infection in shrimp aquaculture may reach 100% within 10 days. VP28 is one of WSSV envelope proteins which is involved in attachment and penetration of virus into shrimp cells. The use of bacteria which expressed VP28 protein as a vaccine is expected to solve WSSV infection in black tiger shrimp (Penaeus monodon). The goal of this experiment is to clone open reading frame (ORF) of VP28 gene into an expression vector pMSP3535VA in Escherichia coli DH5a. pMSP3535VA is a shuttle vector which is able to propagate in E. coli and gram positive bacteria. E. coli clone that contains pMSP3535VA with VP28 ORF inserted was obtained. The recombinant plasmid was characterized using migration analysis, cleavage by restriction enzyme BamHI and EcoRI, Polymerase Chain Reaction (PCR) analysis, and nucleotide sequencing of the inserted DNA. The result of the characterization using migration analysis showed that pMSP3535VA had been ligated with insert DNA, cleavage recombinant plasmid showed that size of insert DNA was 615 base pairs (bps), PCR analysis showed one band with 643 bps in size, and nucleotides of the inserted DNA had 100% homology with VP28 ORF. |
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