| Project/Area Number |
06660413
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Applied veterinary science
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| Research Institution | Nihon University |
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Principal Investigator |
TSUMAGARI Shigehisa College of Bioresource Sciences, Associate Professor., 生物資源科学部, 助教授 (80120509)
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| Project Period (FY) |
1994 – 1995
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| Project Status |
Completed (Fiscal Year 1995)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1995: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1994: ¥1,500,000 (Direct Cost: ¥1,500,000)
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| Keywords | recombinant / bovine / LH / castration / vaccine / antgen / リコンビナ-ト / 牛LH |
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| Research Abstract |
In order to development of recombinant bovine LH vaccine (antigen), I tried to insert bovine LH oligonucleotides into ovalbumin DNA and to synthesize hybrid protein. 1) Four major antigenic regions of ovalbumin were predicted : amino acids 50-72,80-98,250-269 and 300-324.2) The most strongest antigenic and specific site in bovine LH beta subunit was also identified : amino acids 58-71.3) Insertion of bovine LH oligonucleotides to plasmid contained ovalbumin cDNA in pTZ19U was conducted by oligonucleotide-directed mutagenesis using the dut ung method. The sequences were identified by the Sanger dideoxy method. However, defection of a oligonucleotide in the DNA sequences was always found, finally it was predicted a phenomenon in appearance by secondary structure of the fragment. 4) The recombinant protein synthesized from the plasmid was identified as ovalbumin by Western blotting, but was not as bovine LH.5) In order to obtain much more both proteins, the PCR products contained bovine LH and ovalbumin DNA was inserted to pET22b plasmid that is known as E.coli strain one having the strongest performance. The recombinant protein induced from the plasmid was not enough now, furthermore I am proceeding this project.
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