1997 Fiscal Year Final Research Report Summary
Enbancement of freezing tolerance of baker, s yeast by introduction of haedening-inducible genes from Chlorella
| Project/Area Number |
08660164
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
食品科学・栄養科学
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| Research Institution | KYUSHU UNIVERSITY |
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Principal Investigator |
HONJOH Ken-ichi Kyushu Univ., Fac. of Agriculture, Assistant Professor, 農学部, 助手 (00264101)
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| Co-Investigator(Kenkyū-buntansha) |
MIYAMOTO Takahisa Kyushu Univ., Fac. of Agriculture, Assisiate Professor, 農学部, 助教授 (70190816)
HATANO Shoji Kyushu Univ., Fac. of Agriculture, Professor, 農学部, 教授 (30038260)
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| Project Period (FY) |
1996 – 1997
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| Keywords | Chlorella / Yeast / Freezing to tolerance / LEA protein |
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| Research Abstract |
The hiC6 gene, encoding a homologue of a late embryogenesis abundant (LEA) protein, was introduced into Saccharomyces cerevisiae. It was inserted on a multicopy plasmid under the transcriptional control of the yeast GALl promoter. Expression of HIC6 protein was confirmed by immunochemical methods. Expression level of the protein was increased gradually with the induction-time by galactose. With maximum induction time, the freeze-tolerance of yeast transformed with hiC6 gene was approximately 3.3 times (from 20% to 66% survival rate) higher than that of the control yeast.
The isolation of Chlorella genes, which encode glucose 6-phosphate dehydrogenase and omega-3 desaturase, were tried for further enhancement of freezing tolerance of yeast by introduction of them. For PCR amplification of the two genes, oligonucleotide primers were synthesized based on the sequences of the corresponding genes from other organisms. Single strand cDNAs were synthesized using Poly(A)^+RNAs from 6-h hardened Chlorella cells. Parts of coding regions of the genes encoding glucose 6-phosphate dehydrogenase and omega-3 desaturase were amplified by PCR using the single stranded cDNA as templates. The sizes of the amplified fragments were 270 bp and 900 bp, respectively, and confirmed to be parts of the objective genes. Now, we are trying to clone the full-length genes encoding them, using the amplified fhagments as probes.
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