Sub kloning gen hemaglutinin 32 KDA sub unit pili pasteurella multocida serotipe B 2 pada vektor ekspresi eukariot=Sub cloning of the hemagglutinin gene encoding the pasteurella multocida serotype ..
| Main Author: | Perpustakaan UGM, i-lib |
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| Format: | Article NonPeerReviewed |
| Terbitan: |
[Yogyakarta] : Universitas Gadjah Mada
, 2005
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| Subjects: | |
| Online Access: |
https://repository.ugm.ac.id/18019/ http://i-lib.ugm.ac.id/jurnal/download.php?dataId=794 |
Daftar Isi:
- Pasteurella multocida serotypes B:2 is the agent of hemorrhagic septicemia in cattle and buffalo. Controls of the disease were carried out by protecting the bacteria to enter the host. DNA vaccine which cotain of hemagglutinin sub unit pili gene from P.multocida would be expected to induce a good protection. That gene was obtained from pKH405 by digesting the plasmid with EcoR I and Hind III. To have the expression of the gene in animal, the hemagglutinin gene should be cloned into an eukaryotic expression vector. This research had conducted by series activity. Preparation hemagglutinin gene and linear vector were done by digestion pKH405 and pCMV-Script vector used restriction enzyme EcoR I and Hind III, then was been electroeluted. Result of the ligation between the vector and the insert were transformed into competent cells E.coliDH5a with heat shock methods. Isolation of the plasmid used alkaline lysis. The recombinant DNA then was introduced into mice Balb/c with 100 pg and 150 pg doses. Negative controls used the mouse that was injected by PBS. The Detection of expression recombinant DNA passed through measurement of IgG titers antihemagglutinin by ELISA. Titers were defined as resiprocal the highest dilution that had givenirs.JOIll V value. me cut or point value was two time number of OD405 ELISA reader from negative controls (serum from PBS imunization) (Al-Mariri, 2001). The result of this research was gained recombinant DNA between hemagglutinin gene 32 kDa on pCMV-Script vector. Introduction recombinant DNA (pKH408) in mice was not negative reaction after immunization. The recombinant DNA could been detected about its expressions in the expression system of the mice. Keywords : The hemagglutinin gene, P.multocida, Sub Cloning, Expression, Eukaryotic, E.coliDH5a