碩士 / 國立海洋大學 / 生物技術研究所 / 84 / Cold is a major environmental stress for all living organisms.
Poikilothermic animals respond to chronic cold by many cellular
and functional changes in liver tissue. To identify these
changes at molecular level, a new method, differential display
reverse transcription-polymerase chain reaction (DDRT-PCR), was
used to compare the differences of mRNA species expressed
between room temperature and cold temperature. Total RNA,
isolated from 8℃ and 25℃ carp liver cell, were reverse
transcribed into cDNA, and then applied to the polymerase chain
reaction using a combination of (dT)12VA oligomers and 6
arbitary 10mers. Products were analyzed on 6% sequence gel;
bands appearing at 8℃ but not 25℃were recovered, reamplified
by PCR, and subcloned. In 13 subcloned cDNA used as probe for
Northern blot analysis, there were only 4 samples showing
significant expression at 8℃. Comparing these 4 cloned
sequences within gene bank, one reveals 78% identity with
nonmuscle myosin heavy chain. According to these 4 DNA
sequences subcloned, in this study, proceeding 5'RACE to try to
obtain the full length cold-induced genes.
| Identifer | oai:union.ndltd.org:TW/084NTOU0108009 |
| Date | January 1996 |
| Creators | Jen, Chih-Hung, 任志鴻 |
| Contributors | Tang Shye-Jye, 唐世杰 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 60 |
Page generated in 0.0016 seconds