碩士 / 國立高雄海洋科技大學 / 水產食品科學研究所 / 97 / Tilapia (Oreochromis aurea × O. niloticus) was the major commercial aquaculture fish in Taiwan. In order to understand the PCP-induced cytochrome P450 system in tilapia, the full-length cDNA sequence of cytochrome P4501A gene in liver was cloned. And the time course changes of relative expression of CYP1A mRNA in tissues of tilapia were also studied by real-time PCR.
Sequence analysis revealed that full-length CYP1A gene of tilapia (Oreochromis aurea × O. niloticus) was of 2,516 bp consisting of an open reading frame of 1,566 bp encoding putative protein of 521 amino acids, which shows theoretical pI of 6.62 and calculates molecular weight of 59.3 KDa. On a phylogenetic tree inferred from the CYP1A nucleotide sequences, 28 fish species compared here were separated each other to form identical clusters according to each taxonomic level. Tilapia was clustered with the medaka fish and Liza fish, with bootstrap value were 83.9~84.2 and 79.2~80.6% respectively.
In the time course study of relative expression of CYP1A mRNA, after PCP injection, intestine showed a peak induction of 64.2± 5.4 at 5 th day, gills and spleen showed peak induction of 41.7 ± 5.6 and 43.4.7 ± 6.4 at 7th day, liver should a peak induction of 107 ± 17.9 at 14 th day. So, the greatest relative expression of CYP1A mRNA was found in liver, and than intestine, spleen and gills, kidney were decreased in the order. However, the relative expression of CYP1A mRNA of other tissues (brain, muscle, skin, heart, stomach) were all less than 5. By using western blotting, only liver and gills could be detected, other tissues were all not detected. The cause was considered to be the different sensitively of CYP1A antibody for different tissues or to be influence by small quantities of sample. This phenomenon will be studied in further experiment.
| Identifer | oai:union.ndltd.org:TW/097NKIM8084006 |
| Date | January 2009 |
| Creators | Shang-jin Lee, 李尚晉 |
| Contributors | Juang-Lin Lian, 連壯林 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 102 |
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