| Project/Area Number |
62570127
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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 |
Pathological medical chemistry
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| Research Institution | Yamanashi Medical College |
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Principal Investigator |
YOSHIIE OKADA Yamanashi Medical College, 医学部, 助教授 (70037274)
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| Project Period (FY) |
1987 – 1988
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| Project Status |
Completed (Fiscal Year 1988)
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| Budget Amount *help |
¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 1988: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1987: ¥1,000,000 (Direct Cost: ¥1,000,000)
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| Keywords | Esterase D / Retinoblastoma / Human erythrocytes / Molecular diagnosis / Restriction fragment length polymorphism / cDNA cloning / _<gt> 11 / 染色体13番 / エステラーゼ D / λgt11 / 制限酵素切断断片多型 / λgtII |
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| Research Abstract |
1. Esterases D-1 and D-2 were highly purified from human erythrocytes and were characterized.
2. White rabbits were immunized with the purified esterase D-1. The antibodies to esterase D-1 reacted specifically to esterases D.
3. The purified esterase D-1 was digested with proteolytic enzymes. The resulting peptides were separated by high performance liquid chromatography. The primary structures of nine purified peptides were analyzed.
4. cDNA library was prepared from human placenta and was screened with the antibody against esterase D-1. one positive clone was isolated.
5. The insert corresponding to cDNA, approximatelly 300 base paris, was subcloned into pUC 9.
6. The base sequence of the cDNA was determined by the dideoxy method. It encodes the amino acid sequence determined in 3.
7. The original cDNA library prepared from human placenta was rescreened with the cDNA as a probe. The cDNA clone with 1,200 base pairs as an insert was obtained.
8. The base sequence of the longer cDNA showed no methionine codon at the 5'-terminal portion, suggesting that the longer cDNA lacks the 5'-terminal portion encoding approximate 10 amino acids at the amino terminal.
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